Methods of treating chronic pain

ABSTRACT

The invention relates to an anti-CGRP antibody for use in the prevention and/or treatment of chronic pain and/or symptoms of chronic pain, and to a method of treating and/or preventing chronic pain and/or symptoms of chronic pain using an anti-CGRP antibody.

This application is a continuation of U.S. patent application Ser. No.13/892,121 filed on May 10, 2013, now abandoned, which is a continuationof U.S. patent application Ser. No. 13/623,206 filed on Sep. 20, 2012,now abandoned, which is a divisional of U.S. patent application Ser. No.12/920,621 (now U.S. Pat. No. 8,293,239) filed on Sep. 2, 2010, which isa National Stage Application under 35 U.S.C. §371 of PCT/IB2009/050852,filed on Mar. 3, 2009, which claims the benefit of U.S. PatentApplication No. 61/033,558, filed on Mar. 4, 2008.

REFERENCE TO SEQUENCE LISTING

The instant application contains a Sequence Listing which has beensubmitted in ASCII format via EFS-Web and is hereby incorporated byreference in its entirety. The sequence listing contains no new matter.Said ASCII copy, created on Dec. 11, 2015, is named 44306702303USL.txtand is 28,591 bytes in size.

FIELD OF THE INVENTION

The invention relates to an anti-CGRP antibody for use in the preventionand/or treatment of chronic pain and/or symptoms of chronic pain, and toa method of treating and/or preventing chronic pain and/or symptoms ofchronic pain using an anti-CGRP antibody.

BACKGROUND OF THE INVENTION

Chronic pain is a long lasting pain that persists longer than thetemporal course of natural healing of the underlying causative injury ordisease. It serves no beneficial or protective function and an estimated2.7 million people in the UK are invalided due to chronic painconditions.

Cancer pain is one of the most common types of chronic pain anddemonstrates nociceptive components due to tumour growth and neuropathiccomponents due to tumour induced nerve damage. It further involvesstructural damage, nerve entrapment and damage, inflammatory processeswhich lead to the disruption of normal tissue metabolism, the productionof inflammatory prostaglandins and cytokines, and tissue damage.

To date, the main analgesics employed for treatment of chronic pain areopiates and non-steroidal anti-inflammatory drugs (NSAIDS). Both classesof drugs can produce severe side-effects; NSAIDS can cause gastriculceration and renal damage, opiates can cause nausea, constipation,confusion and dependency problems. Opioids fail to produce pain reliefin all individuals suffering chronic pain, even at high doses anddevelopment of analgesic resistance to opioids complicates their utilityfor long term therapy. In particular cancer pain treatment requires theuse of unacceptably high levels of opiates bringing with it side-effectsand at least 20% of treated patients still have uncontrolled pain.

Accordingly, there is a critical medical need to identify newpharmaceutically active compounds that interfere with key steps of thechronic pain process and particularly for the treatment and/orprevention of chronic nociceptive pain and/or symptoms of chronicnociceptive pain.

Surprisingly we have found that administration of an anti-CGRP antibodyis effective, with a peripheral site of action, in the prevention and/ortreatment of chronic pain and in particular chronic nociceptive painsuch as cancer pain.

CGRP (calcitonin gene-related peptide) is a 37 amino acid neuropeptidewhich acts as a neurotransmitter in the central nervous system. It bindswith high affinity to the CGRP receptor, Calcitonin receptor-likereceptor (CRLR), activating adenylate cyclase and protein kinase Aproduction.

Centrally penetrating spinally administered, small molecule selectiveCGRP antagonists have been shown to be useful in the treatment ofneuropathic and nociceptive pain conditions (Adwanikar et al, Pain 2007)suggesting that removal of endogenous CGRP in the spinal cord has anantinociceptive effect. Additionally intrathecal administration ofantiserum against CGRP has been shown to reduce nociceptive behaviour inrodent models of arthritis (Kuraishi, Y., et. al Neurosci. lett (1998)92, 325-329).

Surprisingly we have found that administration of an anti-CGRP antibodyis effective, with a peripheral site of action, in the prevention and/ortreatment of chronic pain and in particular chronic nociceptive painwhen administered peripherally. This peripheral administration routeprovides a distinct advantage over the requirement to administerantibodies intrathecally or spinally, a more high risk and inconvenientprocedure.

BRIEF DESCRIPTION OF THE INVENTION

The present invention provides the use of an anti-CGRP antagonistantibody for the manufacture of a medicament for the prevention and/ortreatment of chronic pain and/or symptoms of chronic pain, wherein themedicament is prepared to be peripherally administered.

The present invention further provides a method of prevention and/ortreatment of chronic pain and/or symptoms of chronic pain, in anindividual, which comprises peripherally administering to saidindividual a therapeutically effective amount of an anti-CGRP antagonistantibody.

In one embodiment, the anti-CGRP antagonist antibody acts peripherallyon administration.

DESCRIPTION OF THE FIGURES

FIG. 1. Effect of antibody G2 on mechanical hypersensitivity to 8 gramvon Frey stimuli in a bone cancer pain model. MRMT-1 injected rats weretreated with antibody G2 or vehicle (PBS+0.01% Tween20) at day 9 postsurgery. Groups were healthy throughout the post-operative period at alltimes, shown by increasing post-operative weight gain (data not shown).Data are mean±SEM of 7-9 rats per group. *p<0.05 versus vehicle treatedgroup at each time point.

FIG. 2. Effect of antibody G2 on mechanical hypersensitivity to 15 grvon Frey stimuli in the bone cancer pain model. MRMT-1 injected ratswere treated with G2 or vehicle (PBS+0.01% Tween20) at day 9 postsurgery. Data are mean±SEM of 7-9 rats per group. *p<0.05 versus vehicletreated group at each time point.

FIG. 3. Effect of antibody G2 on ambulation measured by rota rod. Twoend points were explored. The latency to fall as measurements ofcompound induced impairments in motor co-ordination (A), and rota rodscore, as measurements of ambulation evoked pain (B) in the bone cancerpain model. MRMT-1 injected rats were treated with antibody G2 orvehicle (PBS+0.01% Tween20) at day 9 post surgery. Data are mean±SEM of7-9 rats per group. *p<0.05 versus vehicle treated group at each timepoint.

FIG. 4: Binding assay data demonstrating antibody G1 inhibits thebinding of α-CGRP to the CGRP1 receptor.

FIG. 5a : serum level of anti-CGRP concentration (ug/ml) vs time afterIV administration of 10 mg/kg, measured by anti-IgG ELISA.

FIG. 5b : serum level of anti-CGRP concentration (ug/ml) vs time afterIV administration of 10, 30, 100 mg/kg, measured by anti-IgG ELISA.

FIG. 6: Alanine scan using a C-terminal CGRP fragment (CGRP 25-37;residues 25-37 of SEQ ID NO: 15). The change in affinity is expressed infold loss of affinity and which shows that anti-CGRP antibody G1 bindsto the C-terminal region of human α-CGRP.

FIG. 7: Solution competition by Biacore: CGRP, CGRP fragments orpeptides related in sequence to CGRP were used to determine thespecificity of antibody G1.

FIG. 8: CGRP sequences from human, cynomolgus monkey, rat, mouse, dogand rabbit. Non-conserved residues between species are underlined, theepitope of antibody G1 is in bold (SEQ ID NO: 15, 16, 17, 18, and 34-35,respectively, in order of appearance).

FIG. 9: Data showing G1 inhibits neurogenic flare in the skin startingfrom 90 min post-treatment. G1 was administered by intravenousadministration (1 ml/kg). Data are from 6-8 or 13 rats per group.*p=0.05, **p=0.01 vs vehicle (phosphate buffered saline) treated groupat each time point (AVOVA).

Table 1: Kd and IC50 of anti-CGRP antibodies measured at 25° C. againsthuman α-CGRP [muMab7E9=murine precursor of G1. Its K_(D) for ratβ-CGRP=1 nM. RN4901=murine tool, recognising same epitope as G1 butshowed same affinities and selectivity in rats (β-CGRP K_(D)=17 nM);G1=antibody humanized from muMab7E9 (K_(D) for rat□β□CGRP=0.1 nM).]

Table 2: G1 binding affinities as determined by Biacore

DESCRIPTION OF THE INVENTION

General Techniques

The practice of the present invention will employ, unless otherwiseindicated, conventional techniques of molecular biology (includingrecombinant techniques), microbiology, cell biology, biochemistry andimmunology, which are within the skill of the art. Such techniques areexplained fully in the literature, such as, Molecular Cloning: ALaboratory Manual, second edition (Sambrook et al., 1989) Cold SpringHarbor Press; Oligonucleotide Synthesis (M. J. Gait, ed., 1984); Methodsin Molecular Biology, Humana Press; Cell Biology: A Laboratory Notebook(J. E. Cellis, ed., 1998) Academic Press; Animal Cell Culture (R. I.Freshney, ed., 1987); Introduction to Cell and Tissue Culture (J. P.Mather and P. E. Roberts, 1998) Plenum Press; Cell and Tissue Culture:Laboratory Procedures (A. Doyle, J. B. Griffiths, and D. G. Newell,eds., 1993-1998) J. Wiley and Sons; Methods in Enzymology (AcademicPress, Inc.); Handbook of Experimental Immunology (D. M. Weir and C. C.Blackwell, eds.); Gene Transfer Vectors for Mammalian Cells (J. M.Miller and M. P. Calos, eds., 1987); Current Protocols in MolecularBiology (F. M. Ausubel et al., eds., 1987); PCR: The Polymerase ChainReaction, (Mullis et al., eds., 1994); Current Protocols in Immunology(J. E. Coligan et al., eds., 1991); Short Protocols in Molecular Biology(Wiley and Sons, 1999); Immunobiology (C. A. Janeway and P. Travers,1997); Antibodies (P. Finch, 1997); Antibodies: a practical approach (D.Catty., ed., IRL Press, 1988-1989); Monoclonal antibodies: a practicalapproach (P. Shepherd and C. Dean, eds., Oxford University Press, 2000);Using antibodies: a laboratory manual (E. Harlow and D. Lane (ColdSpring Harbor Laboratory Press, 1999); The Antibodies (M. Zanetti and J.D. Capra, eds., Harwood Academic Publishers, 1995); and Cancer:Principles and Practice of Oncology (V. T. DeVita et al., eds., J. B.Lippincott Company, 1993).

DEFINITIONS

An “antibody” is an immunoglobulin molecule capable of specific bindingto a target, such as a carbohydrate, polynucleotide, lipid, polypeptide,etc., through at least one antigen recognition site, located in thevariable region of the immunoglobulin molecule. As used herein, the termencompasses not only intact polyclonal or monoclonal antibodies, butalso fragments thereof (such as Fab, Fab′, F(ab′)₂, Fv, dAb), singlechain antibodies (ScFv), mutants thereof, chimeric antibodies,diabodies, fusion proteins comprising an antibody portion, and any othermodified configuration of the immunoglobulin molecule that comprises anantigen recognition site. An antibody includes an antibody of any class,such as IgG, IgA, or IgM (or sub-class thereof), and the antibody neednot be of any particular class. Depending on the antibody amino acidsequence of the constant domain of its heavy chains, immunoglobulins canbe assigned to different classes. There are five major classes ofimmunoglobulins: IgA, IgD, IgE, IgG, and IgM, and several of these maybe further divided into subclasses (isotypes), e.g., IgG1, IgG2, IgG3,IgG4, IgA1 and IgA2. The heavy-chain constant domains that correspond tothe different classes of immunoglobulins are called alpha, delta,epsilon, gamma, and mu, respectively. The subunit structures andthree-dimensional configurations of different classes of immunoglobulinsare well known

“Fv” is an antibody fragment that contains a completeantigen-recognition and -binding site. In a two-chain Fv species, thisregion consists of a dimer of one heavy and one light chain variabledomain in tight, non-covalent association. In a single-chain Fv species,one heavy and one light chain variable domain can be covalently linkedby a flexible peptide linker such that the light and heavy chains canassociate in a dimeric structure analogous to that in a two-chain Fvspecies. It is in this configuration that the three CDRs of eachvariable domain interact to define an antigen-binding specificity on thesurface of the VH-VL dimer. However, even a single variable domain (orhalf of a Fv comprising only 3 CDRs specific for an antigen) has theability to recognize and bind antigen, although generally at a loweraffinity than the entire binding site.

The Fab fragment also contains the constant domain of the light chainand the first constant domain (CH1) of the heavy chain. Fab′ fragmentsdiffer from Fab fragments by the addition of a few residues at thecarboxy terminus of the heavy chain CH1 domain including one or morecysteines from the antibody hinge regions. A F(ab)2 fragment is abivalent fragment comprising two Fab fragments linked by a disulfidebridge at the hinge region.

An antibody can have one or more binding sites (for combining withantigen). If there is more than one binding site, the binding sites maybe identical to one another or may be different. For instance, anaturally-occurring immunoglobulin has two identical binding sites, asingle chain antibody or Fab fragment has one binding site, while a“bispecific” or “bifunctional” antibody (diabody) has two differentbinding sites, in terms of sequence and/or antigen/epitope recognition.

An “isolated antibody” is an antibody that (1) is not associated withnaturally-associated components, including other naturally-associatedantibodies, that accompany it in its native state, (2) is free of otherproteins from the same species, (3) is expressed by a cell from adifferent species, or (4) does not occur in nature.

A “monoclonal antibody” refers to a homogeneous antibody populationwherein the monoclonal antibody is comprised of amino acids (naturallyoccurring and non-naturally occurring) that are involved in theselective binding of an antigen. A population of monoclonal antibodiesis highly specific, being directed against a single antigenic site. Theterm “monoclonal antibody” encompasses not only intact monoclonalantibodies and full-length monoclonal antibodies, but also fragmentsthereof (such as Fab, Fab′, F(ab′)₂, Fv), single chain (ScFv), mutantsthereof, fusion proteins comprising an antibody portion, and any othermodified configuration of the immunoglobulin molecule that comprises anantigen recognition site of the required specificity and the ability tobind to an antigen. It is not intended to be limited as regards to thesource of the antibody or the manner in which it is made (e.g., byhybridoma, phage selection, recombinant expression, transgenic animals,etc.).

As used herein, “humanized” antibodies refer to forms of non-human (e.g.murine) antibodies that are specific chimeric immunoglobulins,immunoglobulin chains, or fragments thereof (such as Fv, Fab, Fab′,F(ab′)2 or other antigen-binding subsequences of antibodies) thatcontain minimal sequence derived from non-human immunoglobulin. For themost part, humanized antibodies are human immunoglobulins (recipientantibody) in which residues from a complementarity determining region(CDR) of the recipient are replaced by residues from a CDR of anon-human species (donor antibody) such as mouse, rat, or rabbit havingthe desired specificity, affinity, and .biological activity. In someinstances, Fv framework region (FR) residues of the human immunoglobulinare replaced by corresponding non-human residues. Furthermore, thehumanized antibody may comprise residues that are found neither in therecipient antibody nor in the imported CDR or framework sequences, butare included to further refine and optimize antibody performance. Ingeneral, the humanized antibody will comprise substantially all of atleast one, and typically two, variable domains, in which all orsubstantially all of the CDR regions correspond to those of a non-humanimmunoglobulin and all or substantially all of the FR regions are thoseof a human immunoglobulin consensus sequence. The humanized antibodyoptimally also will comprise at least a portion of an immunoglobulinconstant region or domain (Fc), typically that of a humanimmunoglobulin. Antibodies may have Fc regions modified as described inWO 99/58572. Other forms of humanized antibodies have one or more CDRs(one, two, three, four, five, six) which are altered with respect to theoriginal antibody, which are also termed one or more CDRs “derived from”one or more CDRs from the original antibody.

As used herein, “human antibody” means an antibody having an amino acidsequence corresponding to that of an antibody produced by a human and/orhas been made using any of the techniques for making human antibodiesknown in the art or disclosed herein. This definition of a humanantibody includes antibodies comprising at least one human heavy chainpolypeptide or at least one human light chain polypeptide. One suchexample is an antibody comprising murine light chain and human heavychain polypeptides. Human antibodies can be produced using varioustechniques known in the art. In one embodiment, the human antibody isselected from a phage library, where that phage library expresses humanantibodies (Vaughan et al., 1996, Nature Biotechnology, 14:309-314;Sheets et al., 1998, PNAS, (USA) 95:6157-6162; Hoogenboom and Winter,1991, J. Mol. Biol., 227:381; Marks et al., 1991, J. Mol. Biol.,222:581). Human antibodies can also be made by introducing humanimmunoglobulin loci into transgenic animals, e.g., mice in which theendogenous immunoglobulin genes have been partially or completelyinactivated. This approach is described in U.S. Pat. Nos. 5,545,807;5,545,806; 5,569,825; 5,625,126; 5,633,425; and 5,661,016.Alternatively, the human antibody may be prepared by immortalizing humanB lymphocytes that produce an antibody directed against a target antigen(such B lymphocytes may be recovered from an individual or may have beenimmunized in vitro). See, e.g., Cole et al., Monoclonal Antibodies andCancer Therapy, Alan R. Liss, p. 77 (1985); Boerner et al., 1991, J.Immunol., 147 (1):86-95; and U.S. Pat. No. 5,750,373.

A single chain antibody (scFc) is an antibody in which VL and VH regionsare paired to form a monovalent molecule via a synthetic linker thatenables them to be made as a single protein chain (Bird et al Science,242: 423-426 (1988) and Huston et al., Proc. Natl. Acad. Sci. USA,85:5879-5883 (1988)).

Diabodies are bivalent, bispecific antibodies in which VH and VL domainsare expressed on a single polypeptide chain, but using a linker that istoo short to allow for pairing between the two domains on the samechain, thereby forcing the domains to pair with complementary domains ofanother chain and creating two antigen binding sites.

“Chimeric antibodies” refers to those antibodies wherein one portion ofeach of the amino acid sequences of heavy and light chains is homologousto corresponding sequences in antibodies derived from a particularspecies or belonging to a particular class, while the remaining segmentof the chains is homologous to corresponding sequences in another.Typically, in these chimeric antibodies, the variable region of bothlight and heavy chains mimics the variable regions of antibodies derivedfrom one species of mammals, while the constant portions are homologousto the sequences in antibodies derived from another. One clear advantageto such chimeric forms is that, for example, the variable regions canconveniently be derived from presently known sources using readilyavailable hybridomas or B cells from non human host organisms incombination with constant regions derived from, for example, human cellpreparations. While the variable region has the advantage of ease ofpreparation, and the specificity is not affected by its source, theconstant region being human, is less likely to elicit an immune responsefrom a human subject when the antibodies are injected than would theconstant region from a non-human source. However, the definition is notlimited to this particular example.

A “functional Fc region” possesses at least one effector function of anative sequence Fc region. Exemplary “effector functions” include C1qbinding; complement dependent cytotoxicity (CDC); Fc receptor binding;antibody-dependent cell-mediated cytotoxicity (ADCC); phagocytosis;down-regulation of cell surface receptors (e.g. B cell receptor; BCR),etc. Such effector functions generally require the Fc region to becombined with a binding domain (e.g. an antibody variable domain) andcan be assessed using various assays known in the art for evaluatingsuch antibody effector functions.

A “native sequence Fc region” comprises an amino acid sequence identicalto the amino acid sequence of an Fc region found in nature. A “variantFc region” comprises an amino acid sequence which differs from that of anative sequence Fc region by virtue of at least one amino acidmodification, yet retains at least one effector function of the nativesequence Fc region. Preferably, the variant Fc region has at least oneamino acid substitution compared to a native sequence Fc region or tothe Fc region of a parent polypeptide, e.g. from about one to about tenamino acid substitutions, and preferably from about one to about fiveamino acid substitutions in a native sequence Fc region or in the Fcregion of the parent polypeptide. The variant Fc region herein willpreferably possess at least about 80% sequence identity with a nativesequence Fc region and/or with an Fc region of a parent polypeptide, andmost preferably at least about 90% sequence identity therewith, morepreferably at least about 95% sequence identity therewith.

As used herein “antibody-dependent cell-mediated cytotoxicity” and“ADCC” refer to a cell-mediated reaction in which nonspecific cytotoxiccells that express Fc receptors (FcRs) (e.g. natural killer (NK) cells,neutrophils, and macrophages) recognize bound antibody on a target celland subsequently cause lysis of the target cell. ADCC activity of amolecule of interest can be assessed using an in vitro ADCC assay, suchas that described in U.S. Pat. No. 5,500,362 or 5,821,337. Usefuleffector cells for such assays include peripheral blood mononuclearcells (PBMC) and NK cells. Alternatively, or additionally, ADCC activityof the molecule of interest may be assessed in vivo, e.g., in a animalmodel such as that disclosed in Clynes et al., 1998, PNAS (USA),95:652-656.

As used herein, “Fc receptor” and “FcR” describe a receptor that bindsto the Fc region of an antibody. The preferred FcR is a native sequencehuman FcR. Moreover, a preferred FcR is one which binds an IgG antibody(a gamma receptor) and includes receptors of the FcγRI, FcγRII, andFcγRIII subclasses, including allelic variants and alternatively splicedforms of these receptors. FcγRII receptors include FcγRIIA (an“activating receptor”) and FcγRIIB (an “inhibiting receptor”), whichhave similar amino acid sequences that differ primarily in thecytoplasmic domains thereof. FcRs are reviewed in Ravetch and Kinet,1991, Ann. Rev. Immunol., 9:457-92; Capel et al., 1994, Immunomethods,4:25-34; and de Haas et al., 1995, J. Lab. Clin. Med., 126:330-41. “FcR”also includes the neonatal receptor, FcRn, which is responsible for thetransfer of maternal IgGs to the fetus (Guyer et al., 1976, J. Immunol.,117:587; and Kim et al., 1994, J. Immunol., 24:249).

“Complement dependent cytotoxicity” and “CDC” refer to the lysing of atarget in the presence of complement. The complement activation pathwayis initiated by the binding of the first component of the complementsystem (C1q) to a molecule (e.g. an antibody) complexed with a cognateantigen. To assess complement activation, a CDC assay, e.g. as describedin Gazzano-Santoro et al., J. Immunol. Methods, 202:163 (1996), may beperformed.

As used herein, the terms “G1” and “antibody G1” are usedinterchangeably to refer to an antibody produced by the expressionvectors having deposit numbers ATCC-PTA-6867 and ATCC-PTA-6866. Theamino acid sequence of the heavy chain and light chain variable regionsare shown in SEQ ID Nos. 1 and 2. The CDR portions of antibody G1(including Chothia and Kabat CDRs) are diagrammatically depicted in FIG.5 of WO2007/054809, the content of which is herein incorporated byreference in its entirety.

The polynucleotides encoding the heavy and light chain variable regionsare shown in SEQ ID Nos., 9 and 10. The characterization of antibody G1is described in the Examples of WO2007/054809, the entire content ofwhich is herein incorporated by reference. G1 is a humanized monoclonalblocking antibody (IgG2) which blocks binding and activity of theneuropeptide CGRP (a and b) and its effect of neurogenic vasodilatationcaused by CGRP release. G1 is an IgG2Δa monoclonal anti-CGRP antagonistantibody derived from the murine anti-CGRP antagonist antibodyprecursor, denoted muMAb7E9 as identified in a screen using spleen cellsprepared from a mouse immunized with human and rat CGRP that were fusedwith murine plasmacytoma cells. G1 was created by grafting the muMAb 7E9derived CDRs of light and heavy chain into the closest human germ linesequence followed by the introduction of at least 1 mutation into eachCDR and 2 framework mutations in V_(H). Two mutations were introducedinto the Fc domain of G1 to suppress human Fc-receptor activation. G1and muMab7E9 have been shown to recognise the same epitope.

As used herein, the terms “G2” and “antibody G2” are usedinterchangeably to refer to an anti-rat CGRP mouse monoclonal antibodyas described in Wong H C et al. Hybridoma 12:93-106 (1993). The aminoacid sequence of the heavy chain and light chain variable regions areshown in SEQ ID Nos. 19 and 20. The polynucleotides encoding the heavyand light chain variable regions are shown in SEQ ID Nos. 27 and 28. TheCDR portions of antibody G2 are provided in SEQ ID Nos. 21 to 26. G2 hasbeen shown to recognise the same epitope as G1.

As used herein, “immunospecific” binding of antibodies refers to theantigen specific binding interaction that occurs between theantigen-combining site of an antibody and the specific antigenrecognized by that antibody (i.e., the antibody reacts with the proteinin an ELISA or other immunoassay, and does not react detectably withunrelated proteins).

An epitope that “specifically binds”, or “preferentially binds” (usedinterchangeably herein) to an antibody or a polypeptide is a term wellunderstood in the art, and methods to determine such specific orpreferential binding are also well known in the art. A molecule is saidto exhibit “specific binding” or “preferential binding” if it reacts orassociates more frequently, more rapidly, with greater duration and/orwith greater affinity with a particular cell or substance than it doeswith alternative cells or substances. An antibody “specifically binds”or “preferentially binds” to a target if it binds with greater affinity,avidity, more readily, and/or with greater duration than it binds toother substances. It is also understood by reading this definition that,for example, an antibody (or moiety or epitope) that specifically orpreferentially binds to a first target may or may not specifically orpreferentially bind to a second target. As such, “specific binding” or“preferential binding” does not necessarily require (although it caninclude) exclusive binding. Generally, but not necessarily, reference tobinding means preferential binding.

The terms “polypeptide”, “oligopeptide”, “peptide” and “protein” areused interchangeably herein to refer to polymers of amino acids of anylength. The polymer may be linear or branched, it may comprise modifiedamino acids, and it may be interrupted by non-amino acids. The termsalso encompass an amino acid polymer that has been modified naturally orby intervention; for example, disulfide bond formation, glycosylation,lipidation, acetylation, phosphorylation, or any other manipulation ormodification, such as conjugation with a labeling component. Alsoincluded within the definition are, for example, polypeptides containingone or more analogs of an amino acid (including, for example, unnaturalamino acids, etc.), as well as other modifications known in the art. Itis understood that, because the polypeptides of this invention are basedupon an antibody, the polypeptides can occur as single chains orassociated chains.

“Polynucleotide,” or “nucleic acid,” as used interchangeably herein,refer to polymers of nucleotides of any length, and include DNA and RNA.The nucleotides can be deoxyribonucleotides, ribonucleotides, modifiednucleotides or bases, and/or their analogs, or any substrate that can beincorporated into a polymer by DNA or RNA polymerase. A polynucleotidemay comprise modified nucleotides, such as methylated nucleotides andtheir analogs. If present, modification to the nucleotide structure maybe imparted before or after assembly of the polymer. The sequence ofnucleotides may be interrupted by non-nucleotide components. Apolynucleotide may be further modified after polymerization, such as byconjugation with a labeling component. Other types of modificationsinclude, for example, “caps”, substitution of one or more of thenaturally occurring nucleotides with an analog, internucleotidemodifications such as, for example, those with uncharged linkages (e.g.,methyl phosphonates, phosphotriesters, phosphoamidates, cabamates, etc.)and with charged linkages (e.g., phosphorothioates, phosphorodithioates,etc.), those containing pendant moieties, such as, for example, proteins(e.g., nucleases, toxins, antibodies, signal peptides, ply-L-lysine,etc.), those with intercalators (e.g., acridine, psoralen, etc.), thosecontaining chelators (e.g., metals, radioactive metals, boron, oxidativemetals, etc.), those containing alkylators, those with modified linkages(e.g., alpha anomeric nucleic acids, etc.), as well as unmodified formsof the polynucleotide(s). Further, any of the hydroxyl groups ordinarilypresent in the sugars may be replaced, for example, by phosphonategroups, phosphate groups, protected by standard protecting groups, oractivated to prepare additional linkages to additional nucleotides, ormay be conjugated to solid supports. The 5′ and 3′ terminal OH can bephosphorylated or substituted with amines or organic capping groupsmoieties of from 1 to 20 carbon atoms. Other hydroxyls may also bederivatized to standard protecting groups. Polynucleotides can alsocontain analogous forms of ribose or deoxyribose sugars that aregenerally known in the art, including, for example, 2′-O-methyl-,2′-O-allyl, 2′-fluoro- or 2′-azido-ribose, carbocyclic sugar analogs,α-anomeric sugars, epimeric sugars such as arabinose, xyloses orlyxoses, pyranose sugars, furanose sugars, sedoheptuloses, acyclicanalogs and abasic nucleoside analogs such as methyl riboside. One ormore phosphodiester linkages may be replaced by alternative linkinggroups. These alternative linking groups include, but are not limitedto, embodiments wherein phosphate is replaced by P(O)S(“thioate”), P(S)S(“dithioate”), “(O)NR₂ (“amidate”), P(O)R, P(O)OR′, CO or CH₂(“formacetal”), in which each R or R′ is independently H or substitutedor unsubstituted alkyl (1-20 C) optionally containing an ether (—O—)linkage, aryl, alkenyl, cycloalkyl, cycloalkenyl or araldyl. Not alllinkages in a polynucleotide need be identical. The precedingdescription applies to all polynucleotides referred to herein, includingRNA and DNA.

A “variable region” of an antibody refers to the variable region of theantibody light chain or the variable region of the antibody heavy chain,either alone or in combination. The variable regions of the heavy andlight chain each consist of four framework regions (FR) connected bythree complementarity determining regions (CDRs) also known ashypervariable regions. The CDRs in each chain are held together in closeproximity by the FRs and, with the CDRs from the other chain, contributeto the formation of the antigen-binding site of antibodies. There are atleast two techniques for determining CDRs: (1) an approach based oncross-species sequence variability (i.e., Kabat et al. Sequences ofProteins of Immunological Interest, (5th ed., 1991, National Institutesof Health, Bethesda Md.)); and (2) an approach based on crystallographicstudies of antigen-antibody complexes (Chothia et al. (1989) Nature342:877; Al-Lazikani et al (1997) J. Molec. Biol. 273:927-948)). As usedherein, a CDR may refer to CDRs defined by either approach or by acombination of both approaches.

A “constant region” of an antibody refers to the constant region of theantibody light chain or the constant region of the antibody heavy chain,either alone or in combination.

As used herein, an “anti-CGRP antagonist antibody” (interchangeablytermed “anti-CGRP antibody”) refers to an antibody which is able to bindto CGRP and inhibit CGRP biological activity and/or downstreampathway(s). An anti-CGRP antagonist antibody encompasses antibodies thatblock, antagonize, suppress or reduce (including significantly) CGRPbiological activity. For purpose of the present invention, it will beexplicitly understood that the term “anti-CGRP antagonist antibody”encompass all the previously identified terms, titles, and functionalstates and characteristics whereby the CGRP itself, a CGRP biologicalactivity, or the consequences of the biological activity, aresubstantially nullified, decreased, or neutralized in any meaningfuldegree. Examples of anti-CGRP antagonist antibodies are provided herein.

As used herein, “substantially pure” refers to material which is atleast 50% pure (i.e., free from contaminants), more preferably at least90% pure, more preferably at least 95% pure, more preferably at least98% pure, more preferably at least 99% pure.

A “host cell” includes an individual cell or cell culture that can be orhas been a recipient for vector(s) for incorporation of polynucleotideinserts. Host cells include progeny of a single host cell, and theprogeny may not necessarily be completely identical (in morphology or ingenomic DNA complement) to the original parent cell due to natural,accidental, or deliberate mutation. A host cell includes cellstransfected in vivo with a polynucleotide(s) of this invention.

As used herein, “treatment” is an approach for obtaining beneficial ordesired clinical results. For purposes of this invention, beneficial ordesired clinical results include, but are not limited to, one or more ofthe following: improvement or alleviation of any aspect of chronic painand/or symptom of chronic pain. For purposes of this invention,beneficial or desired clinical results include, but are not limited to,one or more of the following: including lessening severity, alleviationof pain and/or a symptom associated with chronic pain.

An “effective amount” of drug, compound, or pharmaceutical compositionis an amount sufficient to effect beneficial or desired resultsincluding clinical results such as alleviation or reduction in painsensation. An effective amount can be administered in one or moreadministrations. For purposes of this invention, an effective amount ofdrug, compound, or pharmaceutical composition is an amount sufficient totreat, ameliorate, reduce the intensity of and/or prevent chronic painor symptom associated with chronic pain. As is understood in theclinical context, an effective amount of a drug, compound, orpharmaceutical composition may or may not be achieved in conjunctionwith another drug, compound, or pharmaceutical composition. Thus, an“effective amount” may be considered in the context of administering oneor more therapeutic agents, and a single agent may be considered to begiven in an effective amount if, in conjunction with one or more otheragents, a desirable result may be or is achieved.

In one embodiment, “prepared for” herein means the medicament is in theform of a dosage unit or the like suitably packaged and/or marked foruse in peripheral administration.

“Reducing incidence” of chronic pain and/or a symptom associated withchronic pain means any of reducing severity (which can include reducingneed for and/or amount of (e.g., exposure to) other drugs and/ortherapies generally used for these conditions), duration, and/orfrequency.

“Ameliorating” chronic pain and/or a symptom associated with chronicpain means a lessening or improvement of one or more symptoms of chronicpain and/or symptoms associated with chronic pain as compared to notadministering an anti-CGRP antagonist antibody. “Ameliorating” alsoincludes shortening or reduction in duration of a symptom.

“Palliating” chronic pain and/or a symptom associated with chronic painmeans lessening the extent of one or more undesirable clinicalmanifestations of chronic pain in an individual or population ofindividuals treated with an anti-CGRP antagonist antibody in accordancewith the invention.

As used therein, “delaying” the development of chronic pain means todefer, hinder, slow, retard, stabilize, and/or postpone progression ofchronic pain and/or a symptom associated with chronic pain. This delaycan be of varying lengths of time, depending on the history of thedisease and/or individuals being treated. As is evident to one skilledin the art, a sufficient or significant delay can, in effect, encompassprevention, in that the individual does not develop chronic pain. Amethod that “delays” development of the symptom is a method that reducesprobability of developing the symptom in a given time frame and/orreduces extent of the symptoms in a given time frame, when compared tonot using the method. Such comparisons are typically based on clinicalstudies, using a statistically significant number of subjects.

A “biological sample” encompasses a variety of sample types obtainedfrom an individual and can be used in a diagnostic or monitoring assay.The definition encompasses blood and other liquid samples of biologicalorigin, solid tissue samples such as a biopsy specimen or tissuecultures or cells derived therefrom, and the progeny thereof. Thedefinition also includes samples that have been manipulated in any wayafter their procurement, such as by treatment with reagents,solubilization, or enrichment for certain components, such as proteinsor polynucleotides, or embedding in a semi-solid or solid matrix forsectioning purposes. The term “biological sample” encompasses a clinicalsample, and also includes cells in culture, cell supernatants, celllysates, serum, plasma, biological fluid, and tissue samples.

An “individual” or “subject” is a vertebrate, preferably a mammal, morepreferably a human. Mammals include, but are not limited to, farmanimals (such as cows), sport animals, pets (such as cats, dogs andhorses), primates, mice and rats.

As used herein, “vector” means a construct, which is capable ofdelivering, and preferably expressing, one or more gene(s) orsequence(s) of interest in a host cell. Examples of vectors include, butare not limited to, viral vectors, naked DNA or RNA expression vectors,plasmid, cosmid or phage vectors, DNA or RNA expression vectorsassociated with cationic condensing agents, DNA or RNA expressionvectors encapsulated in liposomes, and certain eukaryotic cells, such asproducer cells.

As used herein, “expression control sequence” means a nucleic acidsequence that directs transcription of a nucleic acid. An expressioncontrol sequence can be a promoter, such as a constitutive or aninducible promoter, or an enhancer. The expression control sequence isoperably linked to the nucleic acid sequence to be transcribed.

As used herein, “pharmaceutically acceptable carrier” includes anymaterial which, when combined with an active ingredient, allows theingredient to retain biological activity and is non-reactive with thesubject's immune system. Examples include, but are not limited to, anyof the standard pharmaceutical carriers such as a phosphate bufferedsaline solution, water, emulsions such as oil/water emulsion, andvarious types of wetting agents. Preferred diluents for aerosol orparenteral administration are phosphate buffered saline or normal (0.9%)saline. Compositions comprising such carriers are formulated by wellknown conventional methods (see, for example, Remington's PharmaceuticalSciences, 18th edition, A. Gennaro, ed., Mack Publishing Co., Easton,Pa., 1990; and Remington, The Science and Practice of Pharmacy 20th Ed.Mack Publishing, 2000).

The term “peripherally administered” as used herein refers to the routeby which the a substance, medicament and/or anti-CGRP antagonistantibody is to be delivered, in particular it means not centrally, notspinally, not intrathecally, not delivered directly into the CNS. Theterm refers to administration routes other than those immediatelyforgoing and includes via a route which is oral, sublingual, buccal,topical, rectal, via inhalation, transdermal, subcutaneous, intravenous,intra-arterial, intramuscular, intracardiac, intraosseous, intradermal,intraperitoneal, transmucosal, vaginal, intravitreal, intra-articular,peri-articular, local or epicutaneous.

The term “acts peripherally” as used herein refers to the site of actionof a substance, compound, medicament and/or anti-CGRP antagonistantibody said site being within the peripheral nervous system as opposedto the central nervous system, said compound, medicament and/oranti-CGRP antagonist antibody said being limited by inability to crossthe barrier to the CNS and brain when peripherally administered. Theterm “centrally penetrating” refers to the ability of a substance tocross the barrier to the brain or CNS.

The term “K_(off)”, as used herein, is intended to refer to the off rateconstant for dissociation of an antibody from the antibody/antigencomplex.

The term “K_(d)”, as used herein, is intended to refer to thedissociation constant of an antibody-antigen interaction.

The present invention is directed to a medicament for the preventionand/or treatment of chronic pain and/or symptoms of chronic pain andmethods for prevention and/or treatment of chronic pain and/or symptomsof chronic pain in an individual.

In a first aspect, the invention provides the use of an anti-CGRPantagonist antibody for the manufacture of a medicament for theprevention and/or treatment of chronic pain and/or symptoms of chronicpain, wherein the medicament is prepared for peripheral administrationor wherein the medicament is administered peripherally.

In a second aspect, the invention provides an anti-CGRP antagonistantibody for use in the prevention and/or treatment of chronic painand/or symptoms of chronic pain wherein the antibody is prepared forperipheral administration or wherein the antibody is administeredperipherally.

In third aspect, the invention provides the use of an anti-CGRPantagonist antibody for the manufacture of a medicament forameliorating, controlling, reducing incidence of, or delaying thedevelopment or progression of chronic pain and/or symptoms of chronicpain, wherein the medicament is prepared for peripheral administrationor wherein the medicament is administered peripherally.

In a fourth aspect, the invention provides a method of preventing and/ortreating chronic pain and/or symptoms of chronic pain in an individual,comprising peripheral administration to the individual of an effectiveamount of an anti-CGRP antagonist antibody.

In a fifth aspect, the invention provides a method of ameliorating,controlling, reducing incidence of, or delaying the development orprogression of chronic pain and/or symptoms of chronic pain in anindividual, comprising peripheral administration to the individual of aneffective amount of an anti-CGRP antagonist antibody.

According to a preferred embodiment of the present invention theindividual is preferably a mammal, for example a companion animal suchas a horse, cat or dog or a farm animal such as a sheep, cow or pig.Most preferably the mammal is a human.

According to a preferred embodiment of the present invention themedicament and/or anti-CGRP antagonist antibody is prepared for oral,sublingual, buccal, topical, rectal, inhalation, transdermal,subcutaneous, intravenous, intra-arterial, intramuscular, intracardiac,intraosseous, intradermal, intraperitoneal, transmucosal, vaginal,intravitreal, intra-articular, peri-articular, local or epicutaneousadministration.

According to a further preferred embodiment the medicament is preparedfor peripheral administration prior to and/or during and/or after thedevelopment of chronic pain.

In one embodiment, the anti-CGRP antagonist antibody acts peripherallyon administration. In one embodiment, the anti-CGRP antagonist antibodyis not administered centrally, spinally or intrathecally.

According to a preferred embodiment of the present invention the chronicpain comprises one or more of chronic nociceptive pain, chronicneuropathic pain, chronic inflammatory pain, fibromyalgia, breakthroughpain and persistent pain. The chronic pain may comprise one or more ofhyperalgesia, allodynia, central sensitisation, peripheralsensitisation, disinhibition and augmented facilitation.

According to a further preferred embodiment of the present invention thechronic pain is cancer pain, preferably cancer pain arising frommalignancy or from cancer preferably selected from one or more of:adenocarcinoma in glandular tissue, blastoma in embryonic tissue oforgans, carcinoma in epithelial tissue, leukemia in tissues that formblood cells, lymphoma in lymphatic tissue, myeloma in bone marrow,sarcoma in connective or supportive tissue, adrenal cancer, AIDS-relatedlymphoma, anemia, bladder cancer, bone cancer, brain cancer, breastcancer, carcinoid tumours, cervical cancer, chemotherapy, colon cancer,cytopenia, endometrial cancer, esophageal cancer, gastric cancer, headcancer, neck cancer, hepatobiliary cancer, kidney cancer, leukemia,liver cancer, lung cancer, lymphoma, Hodgkin's disease, lymphoma,non-Hodgkin's, nervous system tumours, oral cancer, ovarian cancer,pancreatic cancer, prostate cancer, rectal cancer, skin cancer, stomachcancer, testicular cancer, thyroid cancer, urethral cancer, bone cancer,sarcomas cancer of the connective tissue, cancer of bone tissue, cancerof blood-forming cells, cancer of bone marrow, multiple myeloma,leukaemia, primary or secondary bone cancer, tumours that metastasize tothe bone, tumours infiltrating the nerve and hollow viscus, tumours nearneural structures. Further preferably the cancer pain comprises visceralpain, preferably visceral pain which arises from pancreatic cancerand/or metastases in the abdomen. Further preferably the cancer paincomprises somatic pain, preferably somatic pain due to one or more ofbone cancer, metastasis in the bone, postsurgical pain, sarcomas cancerof the connective tissue, cancer of bone tissue, cancer of blood-formingcells of the bone marrow, multiple myeloma, leukaemia, primary orsecondary bone cancer.

According to a preferred embodiment of the present invention theanti-CGRP antagonist antibody binds to CGRP, more preferably binds toCGRP and inhibits the ability of CGRP to bind to the CGRP receptor.Preferably the anti-CGRP antagonist antibody binds to both human androdent CGRP, preferably human and rat CGRP. More preferably the antibodybinds to human CGRP, further preferably the anti-CGRP antagonistantibody binds to human α-CGRP or to human α-CGRP and/or β-CGRP. Mostpreferably the anti-CGRP antagonist antibody is an antibody thatexhibits any one or more of the following functional characteristics:(a) binds to CGRP; (b) blocks CGRP from binding to its receptor(s); (c)blocks or decreases CGRP receptor activation, including cAMP activation;(d) inhibits, blocks, suppresses or reduces CGRP biological activity,including downstream pathways mediated by CGRP signalling, such asreceptor binding and/or elicitation of a cellular response to CGRP; (e)prevents, ameliorates, or treats any aspect of chronic pain; (f)increases clearance of CGRP; and (g) inhibits (reduces) CGRP synthesis,production or release.

Antibodies of the invention, including G1 and G2, are known to bind CGRPand remove its biological availability for example in the serum thuspreventing CGRP access to the its receptor and downstream cellularresponses and biological effects of CGRP such as flare and vasodilation.

In a further preferred embodiment of the invention the anti-CGRPantagonist antibody binds to a fragment of CGRP, more preferably to afragment of CGRP as well as to the full length CGRP. Preferably, theanti-CGRP antagonist antibody binds to the C-terminal region or fragmentof CRGP. The C-terminal region or fragment of CRGP preferably comprisesamino acids 19-37 or 25-37 or 29-37 or alternatively 30-37, furtheralternatively 31-37 of CGRP. In a further embodiment, the C-terminalregion or fragment of CRGP preferably comprises amino acids 32-37 mostpreferably 33 to 37 of CGRP. Preferably the CGRP is either α-CGRP orβ-CGRP, further preferably human or rodent, further preferably human orrat, more preferably human, further preferably human α-CGRP or β-CGRP,most preferably human α-CGRP.

In a further preferred embodiment of the invention the anti-CGRPantagonist antibody specifically binds to the amino acid sequence GSKAF(SEQ ID NO: 33). Preferably the sequence GSKAF (SEQ ID NO: 33) of CGRPis the epitope to which the anti-CGRP antagonist antibody binds,preferably at position 33 to 37, most preferably the sequence is GXXXFwhere X can be any amino acid, preferably at positions 33 to 37 of CGRP,the ends defined by amino acids G33 and F37 of CGRP.

In one embodiment, the present invention provides an anti-CGRPantagonist antibody which specifically binds to an epitope defined byamino acids G33 to F37 of CGRP. The anti-CGRP antagonist antibody mayspecifically bind to the epitope defined by the amino acid sequenceGSKAF (SEQ ID NO: 33). In one embodiment, the present invention providesthe use of such an antibody in the uses and methods defined in thevarious aspects of the present invention.

In one embodiment, the anti-CGRP antagonist antibody inhibits orprevents activation of the CGRP receptor. Preferably the anti-CGRPantibody has an IC50 of between 0.0001 (0.1 nM) to 500 μM. In somepreferred embodiments, the IC50 is between 0.0001 μM and, or is atabout, any of 250 μM, 100 μM, 50 μM, 10 μM, 1 μM, 500 nM, 250 nM, 100nM, 50 nM, 20 nM, 15 nM, 10 nM, 5 nM, 1 nM, or 0.5 nM as measured in anin vitro binding assay. In some further preferred embodiments, IC50 isless than any of 500 pM, or 100 pM, or 50 pM, as measured in an in vitrobinding assay. In a further more preferred embodiment IC50 is 1.2 nM or31 nM.

In a further preferred embodiment, the anti-CGRP antagonist antibodyused is capable of competing with an antibody herein above described forthe binding of CGRP or to a fragment of CGRP, or to a fragment of CGRPas well as the full length CGRP, preferably to the C-terminal region orfragment of CRGP, preferably the C-terminal region or fragment of CRGPcomprises amino acids 19-37 or 25-37 or 29-37 or alternatively 30-37,further alternatively 31-37 of CGRP. In a further embodiment, theC-terminal region or fragment of CRGP preferably comprises amino acids32-37, most preferably 33 to 37 of CGRP.

In a further preferred embodiment, the anti-CGRP antagonist antibody orantigen binding portion thereof as used in the invention is capable ofcompeting with an anti-CGRP antagonist antibody herein above described,in particular G1 or G2 as herein described, for:

(a) the binding of CGRP or a fragment of CGRP, or a fragment of CGRP aswell as the full length CGRP, preferably the C-terminal region orfragment of CRGP, preferably the C-terminal region or fragment of CRGPcomprising amino acids 19-37 or 25-37 or 29-37 or alternatively 30-37,further alternatively 31-37, preferably amino acids 32-37, mostpreferably 33 to 37 of CGRP, preferably the CGRP is alpha or beta,preferably beta, more preferably rodent or human, most preferably human.(b) the binding of the epitope sequence GSKAF (SEQ ID NO: 33),preferably at amino acid position 33 to 37 of CGRP as defined in (a),more preferably to the sequence GXXXF, where X is any amino acid,preferably GXXXF at amino acid position 33 to 37 of CGRP as defined in(a).(c) the binding as described in (a) or (b) with substantially the sameKd and/or substantially the same K_(off).(d) binding to CGRP and inhibiting/antagonising CGRP biological activityand/or downstream pathway(s), preferably the CGRP is alpha or beta,preferably beta, more preferably rodent or human, most preferably human.

The anti-CGRP antagonist antibody preferably binds to CGRP, region ofCGRP or fragment of CGRP with a binding affinity (K_(d)) of between0.00001 μM (0.01 nM) to 500 μM. In some preferred embodiments, thebinding affinity (Kd) is between 0.00001 μM and, or is at about, any of250 μM, 100 μM, 50 μM, 10 μM, 1 μM, 500 nM, 250 nM, 100 nM, 50 nM, 20nM, 15 nM, 10 nM, 5 nM, 1 nM, 0.5 nM, 1 nM, 0.05 nM, or 0.01 nM asmeasured in an in vitro binding assay. In some further preferredembodiments, binding affinity (Kd) is less than any of 500 pM, or 100pM, 50 pM, or 10 pM, as measured in an in vitro binding assay. In afurther more preferred embodiment binding affinity (Kd) is 0.04 nM or 16nM.

The anti-CGRP antagonist antibody as used in the present invention maybe selected from the group of: monoclonal antibodies, polyclonalantibodies, antibody fragments (e.g., Fab, Fab′, F(ab′)2, Fv, Fc, ScFvetc.), chimeric antibodies, bispecific antibodies, heteroconjugateantibodies, single chain (ScFv) antibodies, mutants thereof, fusionproteins comprising an antibody portion (e.g., a domain antibody),humanized antibodies, and any other modified configuration of theimmunoglobulin molecule that comprises an antigen recognition site ofthe required specificity, including glycosylation variants ofantibodies, amino acid sequence variants of antibodies, and covalentlymodified antibodies. The anti-CGRP antagonist antibody may be murine,rat, human, or any other origin (including chimeric or humanizedantibodies). In some embodiments, the anti-CGRP antagonist antibody maybe humanized but is more preferably human. Preferably the anti-CGRPantagonist antibody is isolated, further preferably it is substantiallypure. Where the anti-CGRP antagonist antibody is an antibody fragmentthe fragment preferably retains the functional characteristics of theoriginal antibody i.e. the CGRP binding and/or antagonist activity asdescribed in the functional characteristics above.

Examples of anti-CGRP antagonist antibodies are known in the art. Henceaccording to a preferred embodiment of the present invention theanti-CGRP antagonist antibody as used in the present invention ispreferably an anti-CGRP antibody as generally or specifically disclosedin any of (i) WO2007/054809, (ii) WO2007/076336, (iii) Tan et al., Clin.Sci. (Lond). 89:565-73, 1995, (iv) Sigma (Missouri, US), product numberC7113 (clone #4901), (v) Plourde et al., Peptides 14:1225-1229, 1993 orwhich comprises or consists of:

(a) a fragment of said antibody (e.g., Fab, Fab′, F(ab′)2, Fv, Fc, ScFvetc.),

(b) a light chain of said antibody,

(c) a heavy chain of said antibody,

(d) one or more variable region(s) from a light chain and/or a heavychain of said antibody,

(e) one or more CDR(s) (one, two, three, four, five or six CDRs) of saidantibody,

(f) CDR H3 from the heavy chain of said antibody,

(g) CDR L3 from the light chain of said antibody,

(h) three CDRs from the light chain of said antibody,

(i) three CDRs from the heavy chain of said antibody,

(j) three CDRs from the light chain and three CDRs from the heavy chain,of said antibody,

(k) any one or more of (a) through (j).

According to a preferred embodiment of the present invention theanti-CGRP antagonist antibody is antibody G2 or antibody G1. Accordingto a most preferred embodiment of the present the anti-CGRP antagonistantibody used is the anti-CGRP antibody G1 as specifically disclosed inthe patent application WO2007/054809, or comprising its variants shownin Table 6 of WO2007/054809, also including functionally equivalentantibodies to G1, i.e. comprising conservative substitutions of aminoacid residues or one or more deletions or additions of amino acids whichdo not significantly affect their functional characteristics e.g. CRGPbinding or antagonist activity and variants which have enhanced ordecreased activity and/or binding. As used herein, the terms “G1” and“antibody G1” are used interchangeably to refer to an antibody producedby expression vectors having deposit numbers of ATCC PTA-6867 and ATCCPTA-6866 as disclosed in application WO2007/054809.

According to a further embodiment of the present invention, theanti-CGRP antagonist antibody comprises or consists of a polypeptideselected from: (a) antibody G1 or its variants shown in Table 6 ofWO2007/054809; (b) a fragment or a region of antibody G1 or its variantsshown in Table 6 of WO2007/054809; (c) a light chain of antibody G1 orits variants shown in Table 6 of WO2007/054809; (d) a heavy chain ofantibody G1 or its variants shown in Table 6 of WO2007/054809 (e) one ormore variable region(s) from a light chain and/or a heavy chain ofantibody G1 or its variants shown in Table 6 of WO2007/054809; (f) oneor more CDR(s) (one, two, three, four, five or six CDRs) of antibody G1or its variants shown in Table 6 of WO2007/054809; (g) CDR H3 from theheavy chain of antibody G1 or its variants shown in Table 6 ofWO2007/054809; (h) CDR L3 from the light chain of antibody G1 or itsvariants shown in Table 6 of WO2007/054809; (i) three CDRs from thelight chain of antibody G1 or its variants shown in Table 6 ofWO2007/054809; (j) three CDRs from the heavy chain of antibody G1 or itsvariants shown in Table 6 of WO2007/054809; (k) three CDRs from thelight chain and/or three CDRs from the heavy chain, of antibody G1 orits variants shown in Table 6 of WO2007/054809; and (i) an antibodycomprising any one of (b) through (k). The invention also providespolypeptides comprising any one or more of the above. In someembodiments, the at least one, two, three, four, five, or six CDR(s) areat least about 85%, 86%, 87%, 88%, 89%, 90%, 95%, 96%, 97%, 98%, or 99%identical to at least one, two, three, four, five or six CDRs of G1 orits variants shown in Table 6 of WO2007/054809.

Determination of CDR regions is well within the ability of the skilledperson. It is understood that in some embodiments, CDRs can be acombination of the Kabat and Chothia CDR. In some embodiments, the CDRsare the Kabat CDRs. In other embodiments, the CDRs are the Chothia CDRs.

The anti-CGRP antagonist antibody preferably comprises or consists of afragment or a region of the antibody G1 (e.g., Fab, Fab′, F(ab′)2, Fv,Fc, ScFv etc.) or its variants shown in Table 6 of WO2007/054809.Preferably said fragment or region has the functional characteristics ofan anti-CGRP antagonist antibody for example CGRP binding activityand/or antagonist activity and comprises or consists one or more of alight chain, heavy chain, fragment containing one or more variableregions from a light chain and/or a heavy chain, or one or more CDRsfrom a light chain and/or a heavy chain of the antibody G1.

According to a further preferred embodiment of the invention theanti-CGRP antagonist antibody comprises a light chain variable region,LCVR, comprising a peptide with a sequence selected from the groupconsisting of SEQ ID NOs: 28-32 and/or a heavy chain variable region,HCVR, comprising a peptide with a sequence selected from the groupconsisting of SEQ ID NOs: 34-38 of patent application WO2007/076336.

Further preferably the anti-CGRP antagonist antibody comprises an LCVRpolypeptide of a SEQ ID NO as shown in Table 1 of patent applicationWO2007/076336 and further comprises a HCVR polypeptide of a SED ID NO asshown in Table 1 of patent application WO2007/076336.

According to a further embodiment of the invention the anti-CGRPantagonist antibody used comprises a light chain CDR (CDRL) selectedfrom the group consisting of SEQ ID NOs: 8-13 and/or a heavy chain CDR(CDRH) selected from the group consisting of SEQ ID NOs: 14-22 of patentapplication WO2007/076336.

Methods of making and isolating the anti-CGRP antagonist antibodies ofapplication WO2007/076336 and data demonstrating the CGRP binding andantagonist characterisation of the same are described in applicationWO2007/076336.

Preferably the anti-CGRP antagonist antibody for use in the presentinvention comprises a VH domain that is at least 85%, at least 86%, atleast 87%, at least 88%, at least 89%, at least 90%, at least 91%, atleast 92%, at least 93%, at least 94%, at least 95%, at least 96%, atleast 97% at least 98%, at least 99% or 100% identical in amino acidsequence to SEQ ID NO: 1 or SEQ ID NO: 19 presented herein.

Preferably the anti-CGRP antagonist antibody comprises a VL domain thatis at least 85%, at least 86%, at least 87%, at least 88%, at least 89%,at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, atleast 95%, at least 96%, at least 97% at least 98%, at least 99% or 100%identical in amino acid sequence to SEQ ID NO: 2 or SEQ ID NO: 20presented herein.

The anti-CGRP antagonist antibody preferably comprises a VH domain and aVL domain that are at least 85%, at least 86%, at least 87%, at least88%, at least 89%, at least 90%, at least 91%, at least 92%, at least93%, at least 94%, at least 95%, at least 96%, at least 97% at least98%, at least 99% or 100% identical in amino acid sequence to SEQ ID NO:1 and 2 respectively or SEQ ID NO: 19 and 20 presented herein,respectively.

Preferably the anti-CGRP antagonist antibody comprises a VH domain thatis at least 90% identical in amino acid sequence to SEQ ID NO: 1 and aVL domain that is at least 90% identical in amino acid sequence to SEQID NO: 2 presented herein.

Alternatively, the anti-CGRP antagonist antibody preferably comprises aVH domain that is at least 90% identical in amino acid sequence to SEQID NO: 19 and a VL domain that is at least 90% identical in amino acidsequence to SEQ ID NO: 20 presented herein.

The anti-CGRP antagonist antibody preferably comprises at least one CDRselected from the group consisting of: (a). CDR H1 as set forth in SEQID NO: 3 or 21; (b). CDR H2 as set forth in SEQ ID NO: 4 or 22; (c). CDRH3 as set forth in SEQ ID NO: 5 or 23; (d). CDR L1 as set forth in SEQID NO: 6 or 24; (e) CDR L2 as set forth in SEQ ID NO: 7 or 25; (f). CDRL3 as set forth in SEQ ID NO: 8 or 26; and (g). variants of CDR L1, CDRL2 and CDR H2 as shown in Table 6 of WO2007/054809.

According to a preferred embodiment of the present invention theanti-CGRP antagonist antibody heavy chain constant region may be fromany types of constant region, such as IgG, IgM, IgD, IgA, and IgE; andany isotypes, such as IgGI, IgG2, IgG3, and IgG4.

Further preferably the anti-CGRP antagonist antibody comprises a heavychain produced by the expression vector with ATCC Accession No.PTA-6867. Further preferably the anti-CGRP antagonist antibody comprisesa light chain produced by the expression vector with ATCC Accession No.PTA-6866. Further preferably the anti-CGRP antagonist antibody isproduced by the expression vectors with ATCC Accession Nos. PTA-6867 andPTA-6866.

Preferably the anti-CGRP antagonist antibody for use in the presentinvention is antibody G1 or antibody G2 defined herein.

According to a further embodiment of the invention, the anti-CGRPantagonist antibody comprises a modified constant region as for exampledescribed in WO2007/054809. Preferably the modified constant region isimmunologically inert, including partially immunologically inert, suchthat it does not trigger complement mediated lysis, does not stimulateantibody-dependent cell mediated cytotoxicity (ADCC), does not activatemicroglia. Preferably the modified constant region is reduced in one ormore of these activities. Most preferably the constant region ismodified as described in Eur. J. Immunol. (1999) 29:2613-2624; PCTApplication No. PCT/GB99/01441; and/or UK Patent Application No.9809951.8. According to a preferred embodiment of the invention theanti-CGRP antagonist antibody comprises a human heavy chain IgG2constant region comprising the following mutations: A330, P331 to S330,S331 (amino acid numbering with reference to the wildtype IgG2sequence). Eur. J. Immunol. (1999) 29:2613-2624.

Methods of making and isolating the anti-CGRP antagonist antibodies ofapplication WO2007/054809 and data demonstrating the CGRP binding andantagonist characterisation of the same are described in applicationWO2007/054809. Sequences of SEQ ID No. 1 to 14 of said application areprovided herein as SEQ ID No. 1 to 14, respectively.

According to a further embodiment of the present invention themedicament is prepared for peripheral administration between once to 7times per week, further preferably between once to four times per month,further preferably between once to six times per 6 month period, furtherpreferably once to twelve times per year. Preferably the medicament isprepared to be peripherally administered in a period selected from: oncedaily, once every two, three, four, five or six days, weekly, once everytwo weeks, once every three weeks, monthly, once every two months, onceevery three months, once every four months, once every five months, onceevery six months, once every seven months, once every eight months, onceevery nine months, once every ten months, once every eleven months oryearly. According to preferred embodiments the medicament is prepared tobe peripherally administered via a route selected from one or more of;orally, sublingually, buccally, topically, rectally, via inhalation,transdermally, subcutaneously, intravenously, intra-arterially orintramuscularly, via intracardiac administration, intraosseously,intradermally, intraperitoneally, transmucosally, vaginally,intravitreally, epicutaneously, intra-articularly, peri-articularly orlocally.

According to a further embodiment of the present invention themedicament is prepared for peripheral administration with an antibodyconcentration of between 0.1 to 200 mg/ml; preferably at about, orbetween 0.1 and about, any one of 0.5, 1, 5, 10, 15 20, 25, 30, 35, 40,45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 110, 120, 130, 140,150, 160, 170, 180, 190 or 200 mg/ml+/−10% error, most preferably at 50mg/ml.

According to a further embodiment of the present invention themedicament is prepared for peripheral administration with an antibodyconcentration of between 0.1 to 200 mg/kg of body weight; preferably atabout, or between 0.1 and about, any one of 0.5, 1, 5, 10, 15 20, 25,30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 110, 120,130, 140, 150, 160, 170, 180, 190 or 200 mg/kg of body weight+/−10%error, most preferably at 10 mg/kg.

According to a preferred embodiment of the present invention theanti-CGRP antagonist antibody has a half life in-vivo of more than anyone of 2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34,36, 38, 40, 42, 44, 46, 48, 50, 52, 54, 56, 58, 60, 62, 64, 66, 68, 70,72, 74, 76, 78, 80, 82, 84, 86, 88, 90, 92, 94, 96, 98, 100, 102, 104,106, 108, 110, 112, 114, 116, 118, 120, 122, 124, 126, 128, 130, 132,134, 136, 138, 140, 142, 144, 146, 148, 150, 152, 154, 156, 158, 160,162, 164, 166, 168, 170, 172, 174, 176, 178, 180, 182, 184, 186, 188,190, 192, 194, 196, 198, 200, 202, 204, 206, 208 or 210 days+/−1 day,further preferably more than any one of 7, 8, 9, 10, 11, or 12 months.

Preferably the anti-CGRP antagonist antibody has a half life in-vivo ofmore than 6 days.

According to a further preferred embodiment of the present invention,the medicament and/or the anti-CGRP antagonist antibody does not produceeffects of central nervous system and/or cognitive impairment.Preferably the medicament and/or the anti-CGRP antagonist antibody doesnot induce any one ore more of the following: amnesia, confusion,depersonalization, hypesthesia, abnormal thinking, trismus, vertigo,akathisia, apathy, ataxia, circumoral paresthesia, CNS stimulation,emotional lability, euphoria, hallucinations, hostility, hyperesthesia,hyperkinesia, hypotonia, incoordination, libido increase, manicreaction, myoclonus, neuralgia, neuropathy, psychosis, seizure, abnormalspeech, stupor, suicidal ideation; dizziness, somnolence, Insomnia,anxiety, tremor, depression or paresthesia. Most preferably themedicament and/or the anti-CGRP antagonist antibody does not induceimpairment of motor coordination or attention.

According to a further embodiment of the present invention themedicament and/or the anti-CGRP antagonist antibody does not producerespiratory, renal or gastro-intestinal impairment.

According to a further embodiment of the present invention themedicament and/or the anti-CGRP antagonist antibody does not produceeffects of physical and/or psychological dependence. Preferably themedicament and/or the anti-CGRP antagonist antibody does not demonstrateaffinity for opiate, benzodiazepine, phencyclidine (PCP), orN-methyl-D-aspartic acid (NMDA) receptors, or CNS stimulant, or produceany sedating or euphoric effect.

In one embodiment, the anti-CGRP antagonist antibody, on administration,ameliorates, controls, reduces incidence of, or delays the developmentor progression of central pain sensation.

In another embodiment the effect of the anti-CGRP antagonist antibody isequal and/or superior to the effects of NSAIDS and/or opiates in thesame models of chronic pain. In one embodiment, the anti-CGRP antagonistantibody is effective in treating refractory pain populations.

According to a further aspect of the present invention there is providedthe use or method according to any other aspect of the invention whereinthe anti-CGRP antagonist antibody is administered separately,sequentially or simultaneously in combination with one or more furtherpharmacologically active compounds or agents, preferably compounds oragents useful for treating chronic pain. Preferably the additionalagent(s) is/are selected from one or more of:

(i) an opioid analgesic, e.g. morphine, heroin, hydromorphone,oxymorphone, levorphanol, levallorphan, methadone, meperidine, fentanyl,cocaine, codeine, dihydrocodeine, oxycodone, hydrocodone, propoxyphene,nalmefene, nalorphine, naloxone, naltrexone, buprenorphine, butorphanol,nalbuphine or pentazocine;(ii) a nonsteroidal antiinflammatory drug (NSAID), e.g. aspirin,diclofenac, diflusinal, etodolac, fenbufen, fenoprofen, flufenisal,flurbiprofen, ibuprofen, indomethacin, ketoprofen, ketorolac,meclofenamic acid, mefenamic acid, nabumetone, naproxen, oxaprozin,phenylbutazone, piroxicam, sulindac, tolmetin or zomepirac,cyclooxygenase-2 (COX-2) inhibitors, celecoxib; rofecoxib; meloxicam;JTE-522; L-745,337; NS398; or a pharmaceutically acceptable saltthereof;(iii) a barbiturate sedative, e.g. amobarbital, aprobarbital,butabarbital, butabital, mephobarbital, metharbital, methohexital,pentobarbital, phenobartital, secobarbital, talbutal, theamylal orthiopental or a pharmaceutically acceptable salt thereof;(iv) a benzodiazepine having a sedative action, e.g. chlordiazepoxide,clorazepate, diazepam, flurazepam, lorazepam, oxazepam, temazepam ortriazolam or a pharmaceutically acceptable salt thereof;(v) an H₁ antagonist having a sedative action, e.g. diphenhydramine,pyrilamine, promethazine, chlorpheniramine or chlorcyclizine or apharmaceutically acceptable salt thereof;(vi) a sedative such as glutethimide, meprobamate, methaqualone ordichloralphenazone or a pharmaceutically acceptable salt thereof;(vii) a skeletal muscle relaxant, e.g. baclofen, carisoprodol,chlorzoxazone, cyclobenzaprine, methocarbamol or orphrenadine or apharmaceutically acceptable salt thereof;(viii) an NMDA receptor antagonist, e.g. dextromethorphan((+)-3-hydroxy-N-methylmorphinan) or its metabolite dextrorphan((+)-3-hydroxy-N-methylmorphinan), ketamine, memantine, pyrroloquinolinequinone or cis-4-(phosphonomethyl)-2-piperidinecarboxylic acid or apharmaceutically acceptable salt thereof;(ix) an alpha-adrenergic, e.g. doxazosin, tamsulosin, clonidine or4-amino-6,7-dimethoxy-2-(5-methanesulfonamido-1,2,3,4-tetrahydroisoquinol-2-yl)-5-(2-pyridyl)quinazoline;(x) a tricyclic antidepressant, e.g. desipramine, imipramine,amytriptiline or nortriptiline;(xi) an anticonvulsant, e.g. carbamazepine or valproate;(xii) a tachykinin (NK) antagonist, particularly an NK-3, NK-2 or NK-1antagonist, e.g.(αR,9R)-7-[3,5-bis(trifluoromethyl)benzyl]-8,9,10,11-tetrahydro-9-methyl-5-(4-methylphenyl)-7H-[1,4]diazocino[2,1-g][1,7]naphthridine-6-13-dione(TAK-637),5-[[(2R,3S)-2-[(1R)-1-[3,5-bis(trifluoromethyl)phenyl]ethoxy-3-(4-fluorophenyl)-4-morpholinyl]methyl]-1,2-dihydro-3H-1,2,4-triazol-3-one(MK-869), lanepitant, dapitant or3-[[2-methoxy-5-(trifluoromethoxy)phenyl]methylamino]-2-phenyl-piperidine(2S,3S);(xiii) a muscarinic antagonist, e.g oxybutin, tolterodine, propiverine,tropsium chloride or darifenacin;(xiv) a COX-2 inhibitor, e.g. celecoxib, rofecoxib or valdecoxib;(xv) a non-selective COX inhibitor (preferably with GI protection), e.g.nitroflurbiprofen (HCT-1026);(xvi) a coal-tar analgesic, in particular paracetamol;(xvii) a neuroleptic such as droperidol;(xviii) a vanilloid receptor agonist (e.g. resinferatoxin) or antagonist(e.g. capsazepine);(xix) a beta-adrenergic such as propranolol;(xx) a local anaesthetic, such as mexiletine;(xxi) a corticosteriod, such as dexamethasone;(xxii) a serotonin receptor agonist or antagonist;(xxiii) a cholinergic (nicotinic) analgesic;(xxiv) Tramadol (trade mark);(xxv) a PDEV inhibitor, such as sildenafil, vardenafil or taladafil;(xxvi) an alpha-2-delta ligand such as gabapentin or pregabalin;(xxvii) a canabinoid; and(xxviii) an antidepressant, such as amitriptyline (Elavil), trazodone(Desyrel), and imipramine (Tofranil) or anticonvulsants such asphenytoin (Dilantin) or carbamazepine (Tegretol).

According to a further aspect of the present invention there is provideda pharmaceutical composition for the prevention and/or treatment ofchronic pain and/or symptoms of chronic pain or for ameliorating,controlling, reducing incidence of, or delaying the development orprogression of chronic pain and/or symptoms of chronic pain in anindividual, comprising an anti-CGRP antagonist antibody and apharmaceutically acceptable carrier and/or an excipient, wherein thecomposition is prepared to be peripherally administered.

According to a further aspect of the present invention there is provideda kit comprising:

(a) a pharmaceutical composition as defined above; and

(b) instructions for the peripheral administration of an effectiveamount of said pharmaceutical composition to an individual for theprevention and/or treatment of chronic pain and/or symptoms of chronicpain or for ameliorating, controlling, reducing incidence of, ordelaying the development or progression of chronic pain and/or symptomsof chronic pain.

The kit may include one or more containers containing an anti-CGRPantagonist antibody or polypeptide described herein and instructions foruse in accordance with any of the methods and uses of the invention. Thekit may further comprise a description of selecting an individualsuitable for treatment based on identifying whether that individual haschronic pain or is at risk of having chronic pain. The instructions forthe peripheral administration of the pharmaceutical composition mayinclude information as to dosage, dosing schedule and routes ofadministration for the intended treatment.

Preferred features of each aspect of the invention apply equally to eachother aspect mutatis mutandis.

EXAMPLES

The present invention is now described with reference to the followingExamples which are intended to illustrate but not to limit theinvention.

The following examples and figures are made with reference to antibodyG1, an anti-human CGRP human monoclonal antibody; and to antibody G2, ananti-rat CGRP mouse monoclonal antibody (Wong H C et al. Hybridoma12:93-106 (1993)).

Example 1 Set Up of the Rodent Cancer Mechanistic Pain Model

Tumour cells used are syngeneic MRMT-1 rat mammary gland carcinoma cellsdonated from the Novartis Institute (London). The cells are cultured inRPMI 1640 (Gibco) with 10% foetal bovine serum (FCS), 1% L-glutamine 2%penicillin/streptomycin (Gibco). Two brief washes are carried out with0.1% w/v trypsin to release those cells which adhere to the flask, andthen quenched with an equal volume of 10% FCS, followed bycentrifugation of the solution for 3 minutes at 1200 rpm. The pellet iswashed and resuspended in Hanks medium, and the concentration of cellscalculated using a Haemocytometer, with trypan blue staining todetermine the number of dead MRMT-1 cells. The final concentration of3×10³ cells was then obtained by diluting the solution according to thenumber of cells seen. The final solution was kept on ice until time ofinjection.

Male Sprague-Dawley rats weighing close to 170 g at time of surgery wereused to generate the cancer model. Anaesthesia was induced in the ratsusing halothane or isoflurane (1.5-2%) 66% N₂O and 33% O₂, the leg wasshaved over the appropriate area and disinfected with chlorhexidine(Animalcare Ltd, UK.). A small incision in order to expose theanterior-medial surface of the distal end of the tibia was made. A holewas bored in the periosteum using a 0.7 mm dental drill, through which a2 cm polythene tubing was fed 1 cm into the intra-medullar cavity of thetibia. Using a Hamilton syringe the pre-prepared 10 μl of 3×10³ MRMT-1cells were injected through the tubing into the cavity. The tubing wasthen removed and the hole plugged using bone restorative material (IRM,Dentsply USA). The wound was then irrigated with 0.9% saline and closedwith a metal clip. The sham animals were operated upon using the sameprocedure but injected with 10 μl of Hank's solution alone. The animalswere placed in a thermoregulated recovery box until such time that theywere able to be placed back in their housing cages.

Example 2 Assessment of Rodent Anti-CGRP Antibody G2 in the Cancer PainModel

Testing behaviour towards mechanical stimuli uses von Frey filaments(North Coast Medical Inc., USA) to the plantar surface of both theipsilateral and contralateral hindpaw. The rats were placed in a Perspexcubicle with a mesh floor and allowed to acclimatise for 10 minutes.Each von Frey was applied 10 times to each hindpaw alternating betweenthe ipsilateral and contralateral, for duration of 2-3 seconds eachtime. Von Frey Filaments used have bending forces of 1, 5, 9 and 15 g,and a period of 5 minutes was left between ascending von Frey forces. Anocifensive response (a lift) is defined as a brisk withdrawal of thehindpaw and the number of lifts for each paw at each von Frey arerecorded (maximum of 10) and expressed as a percentage response.

An assessment of the efficacy of rodent anti-CGRP antibody G2 inattenuating hypersensitivity to a wide variety of static mechanical,cooling and integrated stimuli was carried out as well as the backgroundbasal pain behaviours in this validated model of cancer induced bonepain. The responses measured are attenuated by standard analgesictreatments such as morphine and gabapentin. All measures were made bythe same scientist in a blind fashion-blinded to the identity of thecompound/control and to the treatment of the animal.

G2-was given IV at 10 mg/kg at day 9 and rats were tested at 2 hrs andthen on days 10, 11, 12 and then 14-18 days post treatment (FIG. 1).

G2 had marked effect on the behavioural responses to the higherintensity mechanical stimuli. Withdrawal frequencies to von Frey 8 gwere reduced two hours after injection and were significantly reducedover that seen in the vehicle treated group on days 11 and 12 (days 2and 3 after injection, p=0.0164 and 0.0311, respectively). In fact, theG2 treated animals now had pain scores similar to the baseline values.By day 14 (day 5 post-G2 injection) there was no discernible differencebetween the G2 and vehicle-treated groups. Both groups reached a similarlevel of hypersensitivity to von Frey 8 g by day 18 after MRMT-1injection (day 9 post-treatment).

A similar reversal in hypersensitivity to von Frey 15 g was alsoapparent. A reduction in hypersensitivity to von Frey 15 g from vehicletreated group was evident at 2 hours post-injection with significanceseen at 2 and 3 days after drug administration (p=0.02 and 0.03respectively). The reductions were lost by 6 days after G2administration and both groups now reached similar maximal withdrawalfrequencies by 18 days post MRMT-1 injection (FIG. 2).

The results indicate that G2 reduces noxious pain experienced in themetastatic bone cancer rat model.

Example 3 Rota Rod Test for Motor Impairment

A further end point tested in the bone cancer pain model was ambulation(by rotarod). The test is to obtain a measurement of locomotorimpairment comparing antibody treated with control animals, eachsubjected to the same test under the same conditions. The rota rod testconsists of 4 rotatable drums divided by flanges with a motor-drivendrum accelerated (Ugo Basile, Comerio, VA, Italy). For a given trial, arat is placed on the rotating rod and the rotation speed is acceleratedfrom 4 to 16 rpm in 2 min. The time of maximal performance is typicallyset at 120 sec. Each animal generally receives three trials per day, at1 hr intervals, for several consecutive days post surgery. The latencyto fall off the rod is represented as mean of the three trials.

No differences were found between the antibody G2 group and the vehiclegroup in the latency to fall from the rotarod during forced ambulation(FIG. 3). This suggests that G2 does not impair pathways involved inmotor co-ordination, or attention and points to a lack of CNSside-effects produced by the antibody.

Example 4 Binding Assay

A binding assay was performed to measure the IC₅₀ of anti-CGRP antibodyG1 and G2 in blocking human α-CGRP from binding to the CGRP1-receptor inSK-N-MC cells. Dose response curves were plotted and K_(i) values weredetermined using the equation: K_(i)=IC₅₀/(1+([ligand]/K_(D)); FIG. 4,where the equilibrium dissociation constant K_(D)=8 pM for human α-CGRPto CGRP1-receptor as present in SK-N-MC cells. The reported IC₅₀ value(in terms of IgG molecules) was converted to binding sites so that itcould be compared with the affinities (K_(D)) determined by Biacore,using N-biotinylated human and rat α-CGRPs were captured on individualflow cells at low levels (typically 100 response units) to provide thereaction surfaces, while an unmodified flow cell served as a referencechannel. G1 was titrated over the chip surface Binding affinities werededuced from the quotient of the kinetic rate constants(K_(D)=k_(off)/k_(on)) see Table 1.

TABLE 1 G2 Mouse Mab 7E9 G1 KD (nM), α-Hu 17 1.0 0.04 IC50 (nM) α-Hu 372.6 1.2 KD (nM) α-Rat 1.0 58 1.2

TABLE 2 N-biotin-CGRP on chip ° C. k_(on) (1/Ms) k_(off) (1/s) T_(1/2)(h) K_(D) (nM) α-human 25 1.85 × 10⁵ 7.80 × 10⁻⁵ 24.68 0.042 α-human 375.87 × 10⁵ 3.63 × 10⁻⁵ 5.30 0.063 β-human 37 4.51 × 10⁵ 6.98 × 10⁻⁵ 2.760.155 α-rat 25 5.08 × 10⁵ 6.18 × 10⁻⁵ 3.12 1.22 α-rat 37 1.55 × 10⁵ 3.99× 10⁻⁴ 0.48 2.57 β-rat 37 5.16 × 10⁵ 7.85 × 10⁻⁵ 2.45 0.152

Binding affinity of G1 for human α and β CGRP was equivalent (Kd=0.155and 0.152 nM respectively). Binding affinity of G2 for rat α and β CGRPwas equivalent (16 and 17 nM, respectively). Additionally G1 bindingaffinity is 40-fold more potent in human than rat for α-CGRP (Kd=0.042and 1.22 nM, respectively) and equi-potent in human and rat for β-CGRP(Kd=0.155 and 0.152 nM, respectively). Antibody G1 also demonstratedgood cross species selectivity and binds rat α-CGRP with the sameaffinity as antibody G2 (around 1.2 nM) Table 2.

G1 binds human and cynomolgus monkey α- and β-CGRP with high affinity(K_(D)=63 and 155 μM, respectively). G1 displays species selectivity forhuman/cyno CGRP and binds α- and β-CGRP from other species e.g. rat withlower affinity (K_(D)=2.57 nM and 152 μM, respectively).

Example 5 Half Life of Anti-CGRP In-Vivo

Serum measurements of anti-CGRP in rat, FIG. 5, indicate that the halflife is of the order of 7 days. The antibody is peripherally restrictedhaving a molecular weight of around 150,000, FIGS. 5a, 5b , i.e. it doesnot cross into the central nervous system or cross the blood brainbarrier.

Example 6 Selectivity of Anti-CGRP Antibody

We determined the specificity of antibody G1 to human or rat CGRP byusing the Biacore chip to “probe” the free concentration of a premixedcomplex of mAb+peptide. As expected when we pre-incubated antibody G1with human or rat CGRP the response was fully blocked. In contrastpre-incubating G1, with an excess of amylin, calcitonin oradrenomedullin was comparable to the control response (G1 plus buffer)demonstrating that G1 did not form a complex with these peptides (FIG.7).

Example 7 Identification of Antibody G1 Binding Epitope

Interaction analysis was conducted at 25° C. on a Biacore 3000™ systemequipped with streptavidin-coated (SA) sensor chips (Biacore AB,Uppsala, Sweden) using a standard Biacore running buffer (HBS-P). Firstwe confirmed that an N-biotinylated 25-37 human α-CGRP fragment boundwith the same affinity to antibody G1, as full-length N-biotinylatedhuman α-CGRP. Each amino acid between position 27-37 was then mutatedindividually to alanine and expressed the fold loss in affinity comparedto the wild-type fragment. N-biotinylated fragments were captured onindividual flow cells at low levels (typically 100 response units) toprovide the reaction surfaces, while an unmodified flow cell served as areference channel. Purified Fab fragments of antibody G1 were generated.Fab fragments were titrated over the chip using 1 μM as the topconcentration of a two-fold dilution series. Association anddissociation phases were monitored at 100 μl/min for 1 minute and 5minutes respectively. Surfaces were regenerated with a mixture of 35%ethanol+25 mM NaOH+0.5M NaCl.

The alanine scan results show that antibody G1 binds to the C-terminalregion of human α-CGRP, particularly residues 25 to 37, and showsspecific binding to a region (i.e. loss of affinity is markedlyincreased when the specific binding region is mutated) which can bedefined as the epitope and which lies within the last 5 C-terminal aminoacids, i.e. from G33A to F37A. Most profound changes in affinity arecaused through the G33A and F37A mutation (FIG. 6). C-terminal Phe isimportant for selectivity of antibody G1 for CGRP vs related peptidesand gene family members (FIG. 8).

Thus, in one embodiment, the present invention provides an anti-CGRPantagonist antibody which specifically binds to an epitope defined byamino acids G33 to F37 of CGRP. The anti-CGRP antagonist antibody mayspecifically bind to the epitope defined by the amino acid sequenceGSKAF (SEQ ID NO: 33), more specifically to the epitope of CGRP isdefined as GXXXF where X can be any amino acid, the G33 and F37 beingthe most important residues of the epitope for defining high affinitybinding of the anti-CGRP antagonist antibody.

Example 8 Analysis of Indicators of Physical or Psychological Dependence

Neither antibody G1 nor antibody G2 demonstrate CNS penetration.Additionally long term observation of animals (rats) dosed with eitherantibody to levels used in the previous examples did not reveal adverseCNS events such as sedation or stimulation/euphoric behaviour incomparison to control animals. These observations indicate an absence ofdependency risk for the antibodies and hence a significantly improvedsafety of the antibodies over current opiates used in current paintherapies.

Example 9 Analysis of Indicators of Gastro-Intestinal Adverse Effects

A 1 month in-vivo rat study with antibody G2 and 1 week comparativestudy with antibody G1 demonstrated that no adverse gastro intestinaleffects were observed on behaviour, food intake, stool production orhistopathology in comparison to control animals. These observationsindicate an absence of gastrointestinal risk for the antibodies andhence a significantly improved safety of the antibodies over currentNSAIDs used in current pain therapies.

Example 10 G1 and G2 as Anti-CGRP Antagonist Antibodies

A known consequence of CGRP biological activity is the generationneurogenic flare when delivered in vivo. G1 and G2 are demonstrated tobe anti-CGRP antagonist antibodies in that they prevent the developmentof neurogenic flare in vivo.

Using a neurogenic skin flare rat model the efficacy of G1 was testedfor its ability to block CGRP effect in vivo. The saphenous nerve in therat is electrically stimulated causing CGRP release from nerve endingsand leading to vasodilation, the resulting changes in blood flow can bemeasured using laser Dopler methods.

Changes in blood flow parameters were expressed as the area under thecurve (AUC, change in arbitrary Doppler flux units multiplied by time).CGRP receptor antagonist CGRP₈₋₃₇ (400 nmol/kg, i.v.) was used as apositive control to validate the specificity of the model (data notshown). To determine the effect of G1 prior to dosing for each animal,the baseline blood flow response to stimulation was established with twosaphenous nerve stimulations each 30 minutes apart. Rats were treatedwith G1 after the blood flow response of the second stimulation hadreturned to baseline levels (approximately 10 minutes post stimulation)and an additional four stimulations at 30 minute intervals wereperformed.

Results (FIG. 9) demonstrated that in vehicle treated animals nosignificant change in blood flow response was but rats treated with G1showed a significant decrease in blood flow response starting at 90 and120 minutes post dose for 10 mg/kg and 1 mg/kg, respectively. Similaractivity was achieved using D2. Additionally in further neurogenic flareand vasodilatation model tests G1 showed marked effect at 7 days post IVdosing (predicted ED₅₀=6 ug/ml in saphenous nerve stimulation model).The conclusions form the tests done is that G1 and G2 demonstrateanti-CGRP antagonist activity.

Similar CGRP function-blocking ability for the antibodies is also shownin the publication, Zeller J, et. al. Br J Pharmacol. 2008 December;155(7):1093-103. Epub 2008 Sep. 8.

The following materials have been deposited with the American TypeCulture Collection, 10801 University Boulevard, Manassas, Va.20110-2209, USA (ATCC):

ATCC Date of Material Antibody No. Accession No. Deposit pDb.CGRP.hKGIG1 heavy chain PTA-6867 Jul. 15, 2005 pEb.CGRP.hFcGl G1 light chainPTA-6866 Jul. 15, 2005

Vector pEb.CGRP.hKGI is a polynucleotide encoding the G1 light chainvariable region and the light chain kappa constant region; and vectorpDb.CGRP.hFcG1 is a polynucleotide encoding the G1 heavy chain variableregion and the heavy chain IgG2 constant region containing the followingmutations: A330P331 to S330S331 (amino acid numbering with reference tothe wildtype IgG2 sequences; see Eur. J. Immunol. (1999) 29:2613-2624).

Below are given antibody sequences useful for practicing the presentinvention.

Antibody sequencesAntibody G1 heavy chain variable region amino acid sequence(SEQ ID NO: 1) EVQLVESGGGLVQPGGSLRLSCAASGFTFSNYWISWVRQAPGKGLEWVAEIRSESDASATHYAEAVKGRFTISRDNAKNSLYLQMNSLRAEDTAVYYCLAYFDYGLAIQNYWGQG TLVTVSSAntibody G1 light chain variable region amino acid sequence(SEQ ID NO: 2)EIVLTQSPATLSLSPGERATLSCKASKRVTTYVSWYQQKPGQAPRLLIYGASNRYLGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCSQSYNYPYTFGQGTKLEIKAntibody G1 CDR H1 (extended CDR) (SEQ ID NO: 3) GFTFSNYWISAntibody G1 CDR H2 (extended CDR) (SEQ ID NO: 4) EIRSESDASATHYAEAVKGAntibody G1 CDR H3 (SEQ ID NO: 5) YFDYGLAIQNY Antibody G1 CDR L1(SEQ ID NO: 6) KASKRVTTYVS Antibody G1 CDR L2 (SEQ ID NO: 7) GASNRYLAntibody G1 CDR L3 (SEQ ID NO: 8) SQSYNYPYTAntibody G1 heavy chain variable region nucleotide sequence(SEQ ID NO: 9) GAAGTTCAGCTGGTTGAATCCGGTGGTGGTCTGGTTCAGCCAGGTGGTTCCCTGCGTCTGTCCTGCGCTGCTTCCGGTTTCACCTTCTCCAACTACTGGATCTCCTGGGTTCGTCAGGCTCCTGGTAAAGGTCTGGAATGGGTTGCTGAAATCCGTTCCGAATCCGACGCGTCCGCTACCCATTACGCTGAAGCTGTTAAAGGTCGTTTCACCATCTCCCGTGACAACGCTAAGAACTCCCTGTACCTGCAGATGAACTCCCTGCGTGCTGAAGACACCGCTGTTTACTACTGCCTGGCTTACTTTGACTACGGTCTGGCTATCCAGAACTACTGGGGTCAGGGTACCCTGGTTACCGTTTCCTCCAntibody G1 light chain variable region nucleotide sequence(SEQ ID NO: 10)GAAATCGTTCTGACCCAGTCCCCGGCTACCCTGTCCCTGTCCCCAGGTGAACGTGCTACCCTGTCCTGCAAAGCTTCCAAACGGGTTACCACCTACGTTTCCTGGTACCAGCAGAAACCCGGTCAGGCTCCTCGTCTGCTGATCTACGGTGCTTCCAACCGTTACCTCGGTATCCCAGCTCGTTTCTCCGGTTCCGGTTCCGGTACCGACTTCACCCTGACCATCTCCTCCCTGGAACCCGAAGACTTCGCTGTTTACTACTGCAGTCAGTCCTACAACTACCCCTACACCTTCGGTCAGGGTACCAAACTGGAAATCAAAAntibody G1 heavy chain full antibody amino acid sequence(including modified IgG2 as described herein) (SEQ ID NO: 11)EVQLVESGGGLVQPGGSLRLSCAASGFTFSNYWISWVRQAPGKGLEWVAEIRSESDASATHYAEAVKGRFTISRDNAKNSLYLQMNSLRAEDTAVYYCLAYFDYGLAIQNYWGQGTLVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSNFGTQTYTCNVDHKPSNTKVDKTVERKCCVECPPCPAPPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTFRVVSVLTVVHQDWLNGKEYKCKVSNKGLPSSIEKTISKTKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPMLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGKAntibody G1 light chain full antibody amino acid sequence(SEQ ID NO: 12)EIVLTQSPATLSLSPGERATLSCKASKRVTTYVSWYQQKPGQAPRLLIYGASNRYLGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCSQSYNYPYTFGQGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGECAntibody G1 heavy chain full antibody nucleotide sequence(including modified IgG2 as described herein) (SEQ ID NO: 13)GAAGTTCAGCTGGTTGAATCCGGTGGTGGTCTGGTTCAGCCAGGTGGTTCCCTGCGTCTGTCCTGCGCTGCTTCCGGTTTCACCTTCTCCAACTACTGGATCTCCTGGGTTCGTCAGGCTCCTGGTAAAGGTCTGGAATGGGTTGCTGAAATCCGTTCCGAATCCGACGCGTCCGCTACCCATTACGCTGAAGCTGTTAAAGGTCGTTTCACCATCTCCCGTGACAACGCTAAGAACTCCCTGTACCTGCAGATGAACTCCCTGCGTGCTGAAGACACCGCTGTTTACTACTGCCTGGCTTACTTTGACTACGGTCTGGCTATCCAGAACTACTGGGGTCAGGGTACCCTGGTTACCGTTTCCTCCGCCTCCACCAAGGGCCCATCTGTCTTCCCACTGGCCCCATGCTCCCGCAGCACCTCCGAGAGCACAGCCGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCAGAACCTGTGACCGTGTCCTGGAACTCTGGCGCTCTGACCAGCGGCGTGCACACCTTCCCAGCTGTCCTGCAGTCCTCAGGTCTCTACTCCCTCAGCAGCGTGGTGACCGTGCCATCCAGCAACTTCGGCACCCAGACCTACACCTGCAACGTAGATCACAAGCCAAGCAACACCAAGGTCGACAAGACCGTGGAGAGAAAGTGTTGTGTGGAGTGTCCACCTTGTCCAGCCCCTCCAGTGGCCGGACCATCCGTGTTCCTGTTCCCTCCAAAGCCAAAGGACACCCTGATGATCTCCAGAACCCCAGAGGTGACCTGTGTGGTGGTGGACGTGTCCCACGAGGACCCAGAGGTGCAGTTCAACTGGTATGTGGACGGAGTGGAGGTGCACAACGCCAAGACCAAGCCAAGAGAGGAGCAGTTCAACTCCACCTTCAGAGTGGTGAGCGTGCTGACCGTGGTGCACCAGGACTGGCTGAACGGAAAGGAGTATAAGTGTAAGGTGTCCAACAAGGGACTGCCATCCAGCATCGAGAAGACCATCTCCAAGACCAAGGGACAGCCAAGAGAGCCACAGGTGTATACCCTGCCCCCATCCAGAGAGGAGATGACCAAGAACCAGGTGTCCCTGACCTGTCTGGTGAAGGGATTCTATCCATCCGACATCGCCGTGGAGTGGGAGTCCAACGGACAGCCAGAGAACAACTATAAGACCACCCCTCCAATGCTGGACTCCGACGGATCCTTCTTCCTGTATTCCAAGCTGACCGTGGACAAGTCCAGATGGCAGCAGGGAAACGTGTTCTCTTGTTCCGTGATGCACGAGGCCCTGCACAACCACTATACCCAGAAGAGCCTGTCCCTGTCTCCAGGAAAGTAAAntibody G1 light chain full antibody nucleotide sequence(SEQ ID NO: 14) GAAATCGTTCTGACCCAGTCCCCGGCTACCCTGTCCCTGTCCCCAGGTGAACGTGCTACCCTGTCCTGCAAAGCTTCCAAACGGGTTACCACCTACGTTTCCTGGTACCAGCAGAAACCCGGTCAGGCTCCTCGTCTGCTGATCTACGGTGCTTCCAACCGTTACCTCGGTATCCCAGCTCGTTTCTCCGGTTCCGGTTCCGGTACCGACTTCACCCTGACCATCTCCTCCCTGGAACCCGAAGACTTCGCTGTTTACTACTGCAGTCAGTCCTACAACTACCCCTACACCTTCGGTCAGGGTACCAAACTGGAAATCAAACGCACTGTGGCTGCACCATCTGTCTTCATCTTCCCTCCATCTGATGAGCAGTTGAAATCCGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCGCGCGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCCGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACCCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGTTCTCCAGTCACAAAGAGCTTCAACCGCGGTGAGTGCTAAAmino acid sequence comparison of human and rat CGRP (human α-CGRP (SEQ IDNO: 15); human β-CGRP (SEQ ID NO: 16); rat α-CGRP (SEQ ID NO: 17); and rat β-CGRP (SEQ ID NO: 18)): (SEQ ID NO: 15)

(SEQ ID NO: 16)

(SEQ ID NO: 17)

(SEQ ID NO: 18)

Antibody G2 heavy chain variable region amino acid sequence(SEQ ID NO: 19)EVQLQQSGPELVKPGASVKMSCKASGYTFTSSVMHWVKQKPGQGLEWIGYINPYNDGTKYNEKFKGKATLTSDKSSSTAYMELSSLTSEDSAVYYCAKGGNDGYWGQGTTLTVS SAntibody G2 light chain variable region amino acid sequence(SEQ ID NO: 20)EIVLTQSPTTMAASPGEKITITCSASSSISSIYLHWYQQKPGFSPKVLIYRASNLASGVPARFSGSGSGTSYSLTIGTMEAEDVATYYCQQGSTIPFTFGSGTKLEIKAntibody G2 CDR H1 (extended CDR) (SEQ ID NO: 21) SSVMHAntibody G2 CDR H2 (extended CDR) (SEQ ID NO: 22) YINPYNDGTKYNEKFKGAntibody G2 CDR H3 (SEQ ID NO: 23) GGNDGY Antibody G2 CDR Ll(SEQ ID NO: 24) SASSSISSIYLH Antibody G2 CDR L2 (SEQ ID NO: 25) RASNLASAntibody G2 CDR L3 (SEQ ID NO: 26) QQGSTIPFTAntibody G2 heavy chain variable region nucleotide sequence(SEQ ID NO: 27)GAGGTCCAGCTGCAGCAGTCTGGACCTGAGCTGGTAAAGCCTGGGGCTTCAGTGAAGATGTCCTGCAAGGCTTCTGGATACACATTCACTAGCTCTGTTATGCACTGGGTGAAGCAGAAGCCTGGGCAGGGCCTTGAGTGGATTGGATATATTAATCCTTACAATGATGGTACTAAGTACAATGAGAAGTTCAAAGGCAAGGCCACACTGACTTCAGACAAATCCTCCAGCACAGCCTACATGGAACTCAGCAGCCTGACCTCTGAGGACTCTGCGGTCTATTACTGTGCAAAAGGGGGTAACGATGGCTACTGGGGCCAAGGCACTACTCTCACAGTCTCCTCAAntibody G2 light chain variable region nucleotide sequence(SEQ ID NO: 28)GAAATTGTGCTCACCCAGTCTCCAACCACCATGGCTGCATCTCCCGGGGAGAAGATCACTATCACCTGTAGTGCCAGCTCAAGTATAAGTTCCATTTACTTGCATTGGTATCAGCAGAAGCCAGGATTCTCCCCTAAAGTCTTGATTTATAGGGCATCCAATCTGGCTTCTGGAGTCCCAGCTCGCTTCAGTGGCAGTGGGTCTGGGACCTCTTACTCTCTCACAATTGGCACCATGGAGGCTGAAGATGTTGCCACTTACTACTGCCAGCAGGGTAGTACTATACCATTCACGTTCGGCTCGGGGACAAAGTTGGAAATAAAAAntibody G2 heavy chain full antibody amino acid sequence(not including Fc domain) (SEQ ID NO: 29)EVQLQQSGPELVKPGASVKMSCKASGYTFTSSVMHWVKQKPGQGLEWIGYINPYNDGTKYNEKFKGKATLTSDKSSSTAYMELSSLTSEDSAVYYCAKGGNDGYWGQGTTLTVSSAKTTPPSVYPLAPGSAAQTNSMVTLGCLVKGYFPEPVTVTWNSGSLSSGVHTFPAVLQSDLYTLSSSVTVPSSTWPSETVTCNVAHPASSTKVDKKIVPRDAntibody G2 light chain full antibody amino acid sequence(SEQ ID NO: 30)EIVLTQSPTTMAASPGEKITITCSASSSISSIYLHWYQQKPGFSPKVLIYRASNLASGVPARFSGSGSGTSYSLTIGTMEAEDVATYYCQQGSTIPFTFGSGTKLEIKRADAAPTVSIFPPSSEQLTSGGASVVCFLNNFYPRDINVKWKIDGSERQNGVLNSWTDQDSKDSTYSMSSTLTLTKDEYERHNSYTCEATHKTSTSPIVKSFNRNECAntibody G2 heavy chain full antibody nucleotide sequence(not including Fc domain) (SEQ ID NO: 31)GAGGTCCAGCTGCAGCAGTCTGGACCTGAGCTGGTAAAGCCTGGGGCTTCAGTGAAGATGTCCTGCAAGGCTTCTGGATACACATTCACTAGCTCTGTTATGCACTGGGTGAAGCAGAAGCCTGGGCAGGGCCTTGAGTGGATTGGATATATTAATCCTTACAATGATGGTACTAAGTACAATGAGAAGTTCAAAGGCAAGGCCACACTGACTTCAGACAAATCCTCCAGCACAGCCTACATGGAACTCAGCAGCCTGACCTCTGAGGACTCTGCGGTCTATTACTGTGCAAAAGGGGGTAACGATGGCTACTGGGGCCAAGGCACTACTCTCACAGTCTCCTCAGCCAAAACGACACCCCCATCTGTCTATCCACTGGCCCCTGGATCTGCTGCCCAAACTAACTCCATGGTGACCCTGGGATGCCTGGTCAAGGGCTATTTCCCTGAGCCAGTGACAGTGACCTGGAACTCTGGATCCCTGTCCAGCGGTGTGCACACCTTCCCAGCTGTCCTGCAGTCTGACCTCTACACTCTGAGCAGCTCAGTGACTGTCCCCTCCAGCACCTGGCCCAGCGAGACCGTCACCTGCAACGTTGCCCACCCGGCCAGCAGCACCAAGGTGGACAAGAAAATTGTGCCCAGGGATAntibody G2 light chain full antibody nucleotide sequence(SEQ ID NO: 32) GAAATTGTGCTCACCCAGTCTCCAACCACCATGGCTGCATCTCCCGGGGAGAAGATCACTATCACCTGTAGTGCCAGCTCAAGTATAAGTTCCATTTACTTGCATTGGTATCAGCAGAAGCCAGGATTCTCCCCTAAAGTCTTGATTTATAGGGCATCCAATCTGGCTTCTGGAGTCCCAGCTCGCTTCAGTGGCAGTGGGTCTGGGACCTCTTACTCTCTCACAATTGGCACCATGGAGGCTGAAGATGTTGCCACTTACTACTGCCAGCAGGGTAGTACTATACCATTCACGTTCGGCTCGGGGACAAAGTTGGAAATAAAACGGGCTGATGCTGCACCAACTGTATCCATCTTCCCACCATCCAGTGAGCAGTTAACATCTGGAGGTGCCTCAGTCGTGTGCTTCTTGAACAACTTCTACCCCAGAGACATCAATGTCAAGTGGAAGATTGATGGCAGTGAACGACAAAATGGTGTCCTGAACAGTTGGACTGATCAGGACAGCAAAGACAGCACCTACAGCATGAGCAGCACCCTCACATTGACCAAGGACGAGTATGAACGACATAACAGCTATACCTGTGAGGCCACTCACAAGACATCAACTTCACCCATCGTCAAGAGCTTCAACAGGAATGAGTGTTAA

The invention claimed is:
 1. A method for reducing incidence of ortreating allodynia in an individual, comprising administering to theindividual an effective amount of an anti-CGRP antagonist antibody,wherein the anti-CGRP antagonist antibody is a monoclonal antibody. 2.The method of claim 1, wherein the anti-CGRP antagonist antibody isperipherally administered.
 3. The method of claim 1, wherein theanti-CGRP antagonist antibody is administered orally, sublingually, viainhalation, transdermally, subcutaneously, intravenously,intra-arterially, intra-articularly, peri-articularly, orintramuscularly.
 4. The method of claim 3, wherein the anti-CGRPantagonist antibody is administered subcutaneously or intravenously. 5.The method of claim 1, wherein the anti-CGRP antagonist antibody actsperipherally on administration.
 6. The method of claim 1, wherein theanti-CGRP antagonist antibody blocks CGRP from binding to its receptor.7. The method of claim 1, wherein the anti-CGRP antagonist antibodyblocks or decreases CGRP receptor activation.
 8. The method of claim 1,wherein the anti-CGRP antagonist antibody increases clearance of CGRP.9. The method of claim 1, wherein the anti-CGRP antagonist antibodyinhibits CGRP synthesis, production or release.
 10. The method of claim1, wherein the anti-CGRP antagonist antibody is a human antibody. 11.The method of claim 1, wherein the anti-CGRP antagonist antibody is ahumanized antibody.
 12. The method of claim 1, wherein the anti-CGRPantagonist antibody binds CGRP with a Kd of 50 nM or less as measured inan in vitro binding assay.
 13. The method according to claim 1, whereinthe anti-CGRP antagonist antibody has a half-life in-vivo of at least 7days.
 14. The method of claim 1, wherein the anti-CGRP antagonistantibody binds a C-terminal fragment having amino acids 25-37 of CGRP ora C-terminal epitope within amino acids 25-37 of CGRP.
 15. The method ofclaim 1, wherein the anti-CGRP antagonist antibody comprises: (a) CDR H1as set forth in SEQ ID NO: 3 or 21; (b) CDR H2 as set forth in SEQ IDNO: 4 or SEQ ID NO: 22; (c) CDR H3 as set forth in SEQ ID NO: 5 or 23;(d) CDR L1 as set forth in SEQ ID NO: 6 or SEQ ID NO: 24; (e) CDR L2 asset forth in SEQ ID NO: 7 or SEQ ID NO: 25; and (f) CDR L3 as set forthin SEQ ID NO: 8 or
 26. 16. The method of claim 1, wherein the anti-CGRPantagonist antibody comprises a V_(H) domain that comprises the aminoacid sequence to SEQ ID NO: 1 and a V_(L) domain that comprises theamino acid sequence to SEQ ID NO:
 2. 17. The method of claim 1, whereinthe anti-CGRP antagonist antibody comprises a heavy chain produced bythe expression vector with ATCC Accession No. PTA-6867.
 18. The methodof claim 1, wherein the anti-CGRP antagonist antibody comprises a lightchain produced by the expression vector with ATCC Accession No.PTA-6866.
 19. The method of claim 1, wherein the individual is a human.20. The method of claim 1, wherein the anti-CGRP antagonist antibody isformulated with a pharmaceutically acceptable excipient.